Compounds and compositions for treatment of cancer

ABSTRACT

Compounds and compositions for treating, preventing or managing cancer are disclosed. The compositions provided herein comprise SNS-595 and N-desmethyl-SNS-595. Also provided are pharmaceutical compositions comprising the compounds and methods of treatment using the compounds and compositions.

1. RELATED APPLICATIONS

This application claims priority to U.S. provisional application Ser.No. 60/812,835, filed Jun. 12, 2006, entitled “Compounds AndCompositions For Treatment Of Cancer”. The disclosure of the abovereferenced application is incorporated by reference herein in itsentirety

2. FIELD

Provided herein are compounds, compositions and methods of their use fortreatment of cancer. Compositions provided herein comprise, for example,(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid and N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid.

3. BACKGROUND

(+)-1,4-Dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid is known for its anti-tumor activity (see, Tsuzuki et al., J. Med.Chem., 47:2097-2106, 2004 and Tomita et al., J. Med. Chem., 45:5564-5575, 2002). Treatment of the following cancers with(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid has been proposed in the literature: bladder cancer, breast cancer,cervical cancer, colon cancer, esophageal cancer, head and neck cancer,liver cancer, lung cancer, melanoma, myeloma, neuroblastoma, ovariancancer, pancreatic cancer, prostate cancer, renal cancer, sarcoma, skincancer, stomach cancer, testicular cancer, thyroid cancer and uterinecancer. Various dosing regimens for the use of this compound have beenreported, for example, see, U.S. Patent Application Pub. Nos.2005-0203120; 2005-0215583 and 2006-0025437 and InternationalApplication No. PCT/US2006/034699, which are incorporated herein byreference in their entirety.

4. SUMMARY

In certain embodiments, provided herein are compositions comprising(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid and N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid. In certain embodiments, the compositions provided herein compriseat least about 97%(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid and from about 0.01% up to about 3% of N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid by total weight of the composition, wherein each of the percentageis based upon total weight of the two components.

In certain embodiments, the compositions are pharmaceuticalcompositions. In certain embodiments, the compositions provided hereincan be synthesized on a process scale. In certain embodiment, thecompositions are useful in the methods of treating, preventing ormanaging one or more cancers in a mammal.

In certain embodiments, provided herein is a compound selected fromO-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid and a glucoranide of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid. In one embodiment, provided herein are pharmaceutical compositionscomprising a compound selected from N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid, O-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid and a glucoranide of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid and methods of using the same for treating, preventing or managingone or more cancers.

In certain embodiments, the methods provided herein compriseadministering N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid in the compositions or dosages described herein to treat, preventand manage one or more cancers.

The types of cancers that can be treated, prevented or managed usingmethods provided herein include, but are not limited to solid tumors andblood born tumors. In certain embodiment, cancer can be cancer of skintissues, organs, blood, and vessels, including, but not limited to,cancers of the bladder, bone or blood, brain, breast, cervix, chest,colon, endrometrium, esophagus, eye, head, kidney, liver, lung, mouth,neck, ovaries, pancreas, prostate, rectum, stomach, testis, throat anduterus. In certain embodiment, the cancer is hematologic malignancy,such as leukemias, lymphomas (Non-Hodgkin's Lymphoma), Hodgkin's disease(also called Hodgkin's Lymphoma) and myeloma. In certain embodiments,the leukemia is chronic lymphocytic leukemia, chronic myeloid leukemia,acute lymphocytic leukemia, acute myelogenous leukemia and acutemyeloblastic leukemia. In certain embodiments, the cancer comprisessolid tumor. In certain embodiments, the cancer can be relapsed,refractory or resistant to conventional therapy. In certain embodiments,the cancer can be metastatic.

Also provided are methods of preparing the compositions and compoundsdescribed herein.

5. BRIEF DESCRIPTION OF FIGURES

FIG. 1 provides results of comparative cell viability assay using(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid (squares), N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid (inverted triangles) and O-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid (triangles).

FIG. 2 provides a scheme for the synthesis of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid.

FIG. 3 provides chromatograms of N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid produced after incubation in liver microsomes (pane A) and aseparate aliquot from the same incubation with authentic N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid standard spiked in after the incubation was terminated (pane B).

FIG. 4 provides an HPLC chromatogram for the reaction products for an invitro reaction of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid with human liver microsomes indicating two desmethyl products,N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid and O-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid. The chromatogram represents a 60 min incubation with 100 μM(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid (major peak indicates product obtained in the reaction with NADPHand minor peak indicates product obtained in the reaction withoutNADPH).

FIG. 5 provides a product ion scan of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid with structures assigned to the major m/z fragments.

FIG. 6 provides a product ion scan of N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid with structures assigned to the major m/z fragments.

FIG. 7 provides an HPLC chromatogram for an exemplary compositioncomprising about 99.2%(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid and about 0.6% N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid.

FIG. 8 shows an x-ray powder diffraction (XRPD) pattern (measured usingCu Kα radiation) for a hydrate of SNS-595.

FIG. 9 shows a differential scanning calorimetry (DSC) and thermalgravimetric analysis (TGA) thermogram for a hydrate of SNS-595.

6. DETAILED DESCRIPTION 6.1 Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of ordinary skillin the art. All patents, applications, published applications and otherpublications noted herein are incorporated by reference in theirentirety. In the event that there are a plurality of definitions for aterm herein, those in this section prevail unless stated otherwise.

As used herein, “SNS-595 refers to(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid. The compound is also known as AG-7352. The chemical structure ofthe compound is provided herein. Unless otherwise designated, SNS-595 or(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid refers to the enantiomerically pure form of the compound.

As used herein, enantiomerically pure(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid is substantially free from(−)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid (i.e., in enantiomeric excess). In other words, the “(+)” form of1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid is substantially free from the “(−)” form of the compound and is,thus, in enantiomeric excess of the “(−)” form. The term“enantiomerically pure” or “pure enantiomer” denotes that the compoundcomprises more than 75% by weight, more than 80% by weight, more than85% by weight, more than 90% by weight, more than 91% by weight, morethan 92% by weight, more than 93% by weight, more than 94% by weight,more than 95% by weight, more than 96% by weight or more than 97% byweight of the enantiomer.

As used herein “N-desmethyl-SNS-595 or “N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid” refers to(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-amino-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid. The chemical structure for the compound is provided elsewhereherein.

As used herein “O-desmethyl-SNS-595 or “O-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid” refers(+)-1,4-dihydro-7-[(3S,4S)-3-hydroxy-4-methylamino-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid. The chemical structure for the compound is provided elsewhereherein.

As used herein, unless specified otherwise, “compound” or “compoundprovided herein” refers to N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-hydroxy-4-methylamino-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid, O-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-hydroxy-4-methylamino-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid or a glucuronide of(+)-1,4-dihydro-7-[(3S,4S)-3-hydroxy-4-methylamino-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid provided herein.

As used herein and unless otherwise indicated, the terms “treat,”“treating” and “treatment” refer to alleviating or reducing the severityof a disease or a symptom associated with the disease or condition beingtreated.

The term “prevention” includes the inhibition of a disease or disorderor a symptom of the particular disease or disorder. In some embodiments,patients with familial history of cancer are candidates for preventiveregimens. Generally, the term “preventing” refers to administration ofthe drug prior to the onset of symptoms, particularly to patients atrisk of cancer.

As used herein and unless otherwise indicated, the term “managing”encompasses preventing the recurrence of the particular disease ordisorder in a patient who had suffered from it, lengthening the time apatient who had suffered from the disease or disorder remains inremission, reducing mortality rates of the patients, and/or maintaininga reduction in severity or avoidance of a symptom associated with thedisease or condition being managed.

As used herein “subject” is an animal, typically a mammal, includinghuman, such as a patient.

As used herein, the term “cancer” includes, but is not limited to, solidtumors and blood born tumors. In certain embodiment, cancer can becancer of skin tissues, organs, blood, and vessels, including, but notlimited to, cancers of the bladder, bone or blood, brain, breast,cervix, chest, colon, endrometrium, esophagus, eye, head, kidney, liver,lung, mouth, neck, ovaries, pancreas, prostate, rectum, stomach, testis,throat and uterus. In certain embodiment, the cancer is hematologicmalignancy, such as leukemias, lymphomas (Non-Hodgkin's Lymphoma),Hodgkin's disease (also called Hodgkin's Lymphoma) and myeloma. Incertain embodiments, the leukemia is chronic lymphocytic leukemia,chronic myeloid leukemia, acute lymphocytic leukemia, acute myelogenousleukemia and acute myeloblastic leukemia. In certain embodiments, thecancer comprises solid tumor. In certain embodiments, the cancer can berelapsed, refractory or resistant to conventional therapy. In certainembodiments, the cancer can be metastatic.

The term “relapsed” refers to a return of cancer cells or symptoms inpatients who have had a previous remission of cancer after therapy.

The terms “refractory” or “resistant” refer to patients that, even aftertreatment or intensive treatment, have residual cancer cells in theirbody.

As used herein, the IC₅₀ refers to an amount, concentration or dosage ofa particular test compound that achieves a 50% inhibition of a maximalresponse in an assay that measures such response.

As used herein, and unless otherwise specified, the terms“therapeutically effective amount” and “effective amount” of a compoundrefer to an amount sufficient to provide a therapeutic benefit in thetreatment, prevention and/or management of a disease, to delay orminimize one or more symptoms associated with the disease or disorder tobe treated. The terms “therapeutically effective amount” and “effectiveamount” can encompass an amount that improves overall therapy, reducesor avoids symptoms or causes of disease or disorder or enhances thetherapeutic efficacy of another therapeutic agent.

As used herein and unless otherwise indicated, the term“pharmaceutically acceptable salt” includes, but is not limited to,salts of acidic or basic groups that can be present in the compoundsprovided herein. Under certain acidic conditions, the compound can forma wide variety of salts with various inorganic and organic acids. Theacids that can be used to prepare pharmaceutically acceptable salts ofsuch basic compounds are those that form salts comprisingpharmacologically acceptable anions including, but not limited to,acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide,calcium edetate, camsylate, carbonate, chloride, bromide, iodide,citrate, dihydrochloride, edetate, edisylate, estolate, esylate,fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate,hexylresorcinate, hydrabamine, hydroxynaphthoate, isethionate, lactate,lactobionate, malate, maleate, mandelate, mesylate, methylsulfate,muscate, napsylate, nitrate, panthothenate, phosphate/diphosphate,polygalacturonate, salicylate, stearate, succinate, sulfate, tannate,tartrate, teoclate, triethiodide and pamoate. Under certain basicconditions, the compound can form base salts with variouspharmacologically acceptable cations. Non-limiting examples of suchsalts include alkali metal or alkaline earth metal salts and,particularly, calcium, magnesium, sodium, lithium, zinc, potassium andiron salts.

As used herein and unless otherwise indicated, the term “hydrate” meansa solid or semi-solid comprising a compound provided herein or a salt ormolecular complex thereof, that further comprises a stoichiometric ornon-stoichiometeric amount of water.

As used herein and unless otherwise indicated, the term “solvate” meansa solid or semi-solid comprising a compound provided herein or a salt ormolecular complex thereof, that further comprises a stoichiometric ornon-stoichiometric amount of one or more types of solvent. The term“solvate” includes hydrates (e.g., hemihydrate, monohydrate,sesquihydrate, dihydrate, trihydrate, tetrahydrate and the like).

The terms “co-administration” and “in combination with” include theadministration of two therapeutic agents (for example, SNS-595 or acomposition provided herein and another anti-cancer agent or secondagent) either simultaneously, concurrently or sequentially with nospecific time limits. In one embodiment, both agents are present in thecell or in the patient's body at the same time or exert their biologicalor therapeutic effect at the same time. In one embodiment, the twotherapeutic agents are in the same composition or unit dosage form. Inanother embodiment, the two therapeutic agents are in separatecompositions or unit dosage forms.

The term “the supportive care agent” refers to any substance thattreats, prevents, manages, reduces or avoids an adverse or unwantedeffect from SNS-595 treatment.

6.2 Compounds and Compositions

SNS-595 is enantiomerically pure(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid, which is also known as AG-7352 or SP-9595. SNS-595 has thefollowing chemical structure:

In one embodiment, provided herein is a hydrate comprising SNS-595. Incertain embodiment, the hydrate exhibits an X-ray powder diffractionpattern comprising a peak at approximately 8.2 degrees 2θ. In anotherembodiment, the hydrate exhibits an X-ray powder diffraction patternfurther comprising peaks at approximately 6.9, 11.1 and 18.8 degrees 2θ.In a further embodiment, the hydrate exhibits an X-ray powderdiffraction pattern further comprising peaks at approximately 16.4,17.5, 20.8 and 24.9 degrees 2θ.

In one embodiment, the hydrate exhibits a differential scanningcalorimetry thermogram comprising an endothermic event with an onsettemperature of approximately 126.5° C. when heated from approximately25° C. to approximately 350° C. at approximately 10° C./min. In anotherembodiment, the hydrate exhibits a differential scanning calorimetrythermogram further comprising an endothermic event with an onsettemperature of approximately 273.3° C. In a further embodiment, thehydrate exhibits a thermogravimetric analysis thermogram comprising aweight loss of approximately 4.4% when heated from approximately 25° C.to approximately 200° C. at approximately 10° C./min.

In certain embodiments, the hydrate comprises between approximately 0.8and 1.2 molar equivalents of water per mole of SNS-595. In anotherembodiment, the hydrate comprises between approximately 0.9 to 1.1 molarequivalents of water per mole of SNS-595. In another embodiment, thehydrate comprises between approximately 0.95 to 1.05 molar equivalentsof water per mole of SNS-595. In another embodiment, the hydratecomprises between approximately 0.98 to 1.02 molar equivalents of waterper mole of SNS-595.

In one embodiment, provided herein are methods of treatment, preventionor amelioration of one or more cancers comprising administeringN-desmethyl-SNS-595 or a pharmaceutically acceptable salt, solvate orhydrate thereof in the compositions and dosages described herein.N-desmethyl SNS-595 has the following chemical structure:

In one embodiment, provided herein is a compound selected fromO-desmethyl-SNS-595 and a glucuronide of SNS-595 and pharmaceuticallyacceptable derivatives, such as salts, solvates or hydrates of thecompound. In one embodiment, provided herein is O-desmethyl-SNS-595 or apharmaceutically acceptable salt, solvate or hydrate thereof and methodsof use thereof for treatment, prevention or amelioration of one or morecancers. O-desmethyl SNS-595 has the following chemical structure:

In one embodiment, provided herein is a glucuronide of SNS-595 or apharmaceutically acceptable salt, solvate or hydrate thereof and methodsof use thereof for treatment, prevention or amelioration of one or morecancers. The glucuronide of SNS-595 has the following chemicalstructure:

In one embodiment, provided herein is a composition comprising SNS-595and N-desmethyl-SNS-595. In certain embodiments, the compositionprovided herein comprises at least 97% SNS-595 by weight of the activecomponent. In one embodiment, the composition comprises at least 98%, atleast 98.3%, at least 98.5%, at least 98.7%, at least 98.9%, at least99%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, atleast 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least99.9%, at least 99.95% or at least 99.99% SNS-595 by weight of theactive component. In certain embodiments, the percentages of SNS-595 andN-desmethyl-SNS-595 are based upon total weight of the two components.

In certain embodiments, provided herein is a composition comprisingSNS-595 and from about 0.01% up to about 3% by weight ofN-desmethyl-SNS-595. In one embodiment, the composition comprises fromabout 0.1% up to about 2.5%, about 0.2% up to about 2.3%, about 0.3% upto about 2%, about 0.4% up to about 2%, about 0.5% up to about 2%, about0.6% up to about 2%, about 0.7% up to about 2% or about 0.8% up to about2% by weight of N-desmethyl-SNS-595. In another embodiment, thecomposition comprises from about 0.3% up to about 1.8%, about 0.4% up toabout 1.5% or about 0.6% up to about 1.5% of N-desmethyl-SNS-595. Inother embodiments, the composition comprises about 0.1%, 0.2%, 0.3%,0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%,1.6%, 1.7%, 1.8%, 1.9% or 2% of N-desmethyl-SNS-595 based upon totalweight of SNS-595 and N-desmethyl-SNS-595.

In certain embodiments, the composition comprises N-desmethyl-SNS-595and from about 99.99% to about 97% of SNS-595 by weight. In oneembodiment, the composition comprises from about 99.95% to about 97.5%,about 99.9% to about 98%, about 99.7% to about 98.5% or about 99.5% toabout 99% of SNS-595 by weight. In another embodiment, the compositioncomprises from about 99.7% to about 98.2% of SNS-595 by weight. In otherembodiments, the composition comprises about 99.9% 99.8%, 99.7%, 99.6%,99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99%, 98.9%, 98.8%, 98.7%, 98.6%,98.5%, 98.4%, 98.3%, 98.2%, 98.1% or 98% of SNS-595 by weight based upontotal weight of SNS-595 and N-desmethyl-SNS-595.

In certain embodiments, provided herein is a composition that comprisesfrom about 99.99% to about 97% of SNS-595 and from about 0.01% up toabout 3% by weight of N-desmethyl-SNS-595, wherein each of thepercentage is based upon total weight of the two components. In anotherembodiment, the composition comprises from about 99.95% to about 98% ofSNS-595 and from about 0.05% up to about 2% by weight ofN-desmethyl-SNS-595. In one embodiment, the composition comprises fromabout 99.5% to about 98% of SNS-595 and from about 0.5% up to about 2%by weight of N-desmethyl-SNS-595. In other embodiments, the compositioncomprises about 99.9% SNS-595 and about 0.1% N-desmethyl-SNS-595, about99.8% SNS-595 and about 0.2% N-desmethyl-SNS-595, about 99.7% SNS-595and about 0.3% N-desmethyl-SNS-595, about 99.6% SNS-595 and about 0.4%N-desmethyl-SNS-595, 99.5% SNS-595 and about 0.5% N-desmethyl-SNS-595,about 99.4% SNS-595 and about 0.6% N-desmethyl-SNS-595, about 99.3%SNS-595 and about 0.7% N-desmethyl-SNS-595, about 99.2% SNS-595 andabout 0.8% N-desmethyl-SNS-595, about 99.1% SNS-595 and about 0.9%N-desmethyl-SNS-595, about 99% SNS-595 and about 1% N-desmethyl-SNS-595,about 98.9% SNS-595 and about 1.1% N-desmethyl-SNS-595, about 98.8%SNS-595 and about 1.2% N-desmethyl-SNS-595, about 98.7% SNS-595 andabout 1.3% N-desmethyl-SNS-595, about 98.6% SNS-595 and about 1.4%N-desmethyl-SNS-595, about 98.5% SNS-595 and about 1.5%N-desmethyl-SNS-595, about 98.4% SNS-595 and about 1.6%N-desmethyl-SNS-595, about 98.3% SNS-595 and about 1.7%N-desmethyl-SNS-595, about 98.2% SNS-595 and about 1.8%N-desmethyl-SNS-595, about 98.1% SNS-595 and about 1.9%N-desmethyl-SNS-595, or about 98% SNS-595 and about 2%N-desmethyl-SNS-595 by weight. Each of the above is by weight of theactive component.

In certain embodiments, provided herein is a composition comprisingabout 97% to about 99.99% by weight of a hydrate of SNS-595 and fromabout 0.01% up to about 3% by weight of an N-desmethyl-SNS-595, whereineach of the percentages is based upon total weight of the twocomponents. In one embodiment, the composition comprises about 0.8% byweight N-desmethyl-SNS-595. In one embodiment, the composition comprisesabout 1.5% by weight N-desmethyl-SNS-595. In one embodiment, thecomposition comprises about 98.5% by weight of the hydrate of SNS-595.In one embodiment, the composition comprises about 99.2% by weight ofthe hydrate of SNS-595.

In certain embodiments, provided herein are methods for assaying theamount of N-desmethyl-SNS-595 in a composition or a biological sample.In certain embodiments, the composition comprises from about 99.99% toabout 97% of SNS-595 and from about 0.01% up to about 3% by weight ofN-desmethyl-SNS-595. Such methods can be accomplished by techniquesknown to one of skill in the art and described herein, such as HPLCassays.

Also provided are pharmaceutical compositions comprising the compoundsand compositions disclosed herein along with a pharmaceuticallyacceptable carrier, excipient or adjuvant. In one embodiment, thepharmaceutical composition comprises a composition comprising SNS-595and N-desmethyl-SNS-595. In another embodiment, the pharmaceuticalcomposition comprises a compound selected from N-desmethyl-SNS-595,O-desmethyl-SNS-595, glucuronide of SNS-595 and pharmaceuticallyacceptable salt thereof. In one embodiment, the pharmaceuticalcomposition comprises a hydrate of SNS-595, N-desmethyl-SNS-595 and apharmaceutically acceptable carrier, excipient or adjuvant.

5.3 Methods of Preparation

The compositions provided herein can be prepared using techniquesdescribed herein and techniques known to one of skill in the art, forexample, Example C-1 of U.S. Pat. No. 5,817,669, titled “Compounds,processes for the preparation thereof and anti-tumor agents,” issuedOct. 6, 1998, and in Japanese Patent Application No. Hei 10-173986, toChikugi et al., which are incorporated herein by reference in theirentirety. Certain exemplary pharmaceutical compositions comprisingSNS-595 and methods of using the same are described in U.S. PatentApplication Pub. Nos. 2005-0203120; 2005-0215583 and 2006-0025437, whichare incorporated herein by reference in their entirety.

In an exemplary method for preparation, compositions provided herein areprepared from 2,6-dichloronicotinic acid and either 2-pyrroline or1,4-dichloro-2-butene as illustrated in the following scheme (Tsuzuki etal., J. Med. Chem., 47:2097-2106, 2004 and Tomita et al., J. Med. Chem.,45: 5564-5575, 2002):

An exemplary synthetic process is described in the Examples section andillustrated in FIG. 2.

5.4 Dosages

In certain representative embodiments, the method of treating,preventing or managing cancers provided herein comprises administeringto a patient on the basis of body surface area, a dose of about 1mg/m²-150 mg/m² or about 1 mg/m²-75 mg/m² of a compound or compositionprovided herein. In another embodiment, the method of comprisesadministering a dose of about 1 mg/m²-60 mg/m² of the compound orcomposition. In another embodiment, the method of comprisesadministering a dose of about 10 mg/m²-60 mg/m² of the compound orcomposition. In another embodiment, the method comprises administering adose of about 1 mg/m²-50 mg/m² of the compound or composition. Inanother embodiment, the method comprises administering a dose of about 1mg/m²-48 mg/m² of the compound or composition. In another embodiment,the method of comprises administering a dose of about 10 mg/m²-48 mg/m²of the compound or composition. In another embodiment, the methodcomprises administering a dose of about 1 mg/m²-24 mg/m² of the compoundor composition. In another embodiment, the method comprisesadministering a dose of about 3 mg/m²-27 mg/m² of the compound orcomposition provided herein on the basis of body surface area.

The weight of the compound or composition in an administered dose isbased on total weight of the active compound or composition,respectively, e.g., total weight of SNS-595 and N-desmethyl SNS-595 ortotal weight of N-desmethyl-SNS-595.

Body surface area calculations can be calculated for example, with theMosteller formula wherein:

BSA(m²)=square root of [(height(cm)×weight(kg)/3600].

In another embodiment, the dose is about 10 mg/m²-60 mg/m² or about 10mg/m²-48 mg/m² of the compound or composition on the basis of bodysurface area. In another embodiment, the dose is about 3 mg/m²-24 mg/m²of the compound or composition on the basis of body surface area. Inanother embodiment, the dose is about 3 mg/m²-18 mg/m² on the basis ofbody surface area. In another embodiment, the dose is about 3 mg/m²-15mg/m² of the compound or composition. In another embodiment, the dose isabout 1 mg/m², 2 mg/m², 3 mg/m², 4 mg/m², 5 mg/m², 6 mg/m², 7 mg/m², 8mg/m², 9 mg/m², 10 mg/m², 11 mg/m², 12 mg/m², 13 mg/m², 14 mg/m², 15mg/m², 16 mg/m², 17 mg/m², 18 mg/m², 19 mg/m², 20 mg/m², 21 mg/m², 22mg/m², 23 mg/m², 24 mg/m², 25 mg/m², 26 mg/m², 27 mg/m², 28 mg/m², 30mg/m², 45 mg/m², 48 mg/m², 60 mg/m² or 75 mg/m² of the compound orcomposition on the basis of body surface area. In another embodiment,the dose is about 40 mg/m², 41 mg/m², 42 mg/m², 43 mg/m², 44 mg/m², 45mg/m², 46 mg/m², 47 mg/m², 48 mg/m², 49 mg/m², 50 mg/m², 51 mg/m², 52mg/m², 53 mg/m², 54 mg/m², 55 mg/m², 56 mg/m², 57 mg/m², 58 mg/m², 59mg/m², 60 mg/m², 62 mg/m², 65 mg/m², 68 mg/m², 70 mg/m², 72 mg/m², 75mg/m², 80 mg/m², 84 mg/m², 85 mg/m² or 90 mg/m² of the compound orcomposition on the basis of body surface area. In another embodiment,the dose is about 3 mg/m², 12 mg/m², 15 mg/m², 18 mg/m², 48 mg/m² or 60mg/m² of the compound or composition. In another embodiment, the dose isabout 15 mg/m² of the compound or composition provided herein on thebasis of body surface area.

The administered dose of the compound or composition provided herein canbe expressed in units other than as mg/m². For example, doses can beexpressed as mg/kg. One of ordinary skill in the art would readily knowhow to convert doses from mg/m² to mg/kg to given either the height orweight of a subject or both (see,www.fda.gov/cder/cancer/animalframe.htm). For example, a dose of 1mg/m²-30 mg/m² for a 65 kg human is approximately equal to 0.026mg/kg-0.79 mg/kg. In another example, a dose of 3 mg/m² for a 65 kghuman is approximately equal to 0.078 mg/kg.

In certain embodiments, the administered dose of the compound orcomposition provided herein can be delivered as a single dose (e.g. asingle bolus IV injection) or over a 24-hour period (e.g., continuousinfusion over time or divided bolus doses over time) and is repeateduntil the patient experiences stable disease or regression or until thepatient experiences disease progression or unacceptable toxicity. Stabledisease or lack thereof is determined by methods known in the art, suchas evaluation of patient symptoms, physical examination and othercommonly accepted evaluation modalities.

The amount of the pharmaceutical composition administered according tothe methods provided herein will depend on various factors, such as themammal being treated, the severity of the disorder or symptom of thedisorder, the active ingredient present, the manner of administration,the frequency of administration and the judgment of the prescribingphysician. The amount can be empirically determined by the physician.

In some embodiments, the frequency of administration is in the range ofabout a daily dose to about a monthly dose. In certain embodiments,administration is once a day to once every other day or from twice aweek, once every week, once every two weeks, once every three weeks oronce every four weeks. In one embodiment, the pharmaceutical compositionprovided herein is administered once a week.

In certain embodiments, the compound or composition provided herein iscyclically administered to a patient. Cycling therapy involves theadministration of an the compound or composition for a period of time,followed by a rest for a period of time, and repeating this sequentialadministration. Cycling therapy can reduce the development of resistanceto one or more of the therapies, avoid or reduce the side effects of oneof the therapies, and/or improve the efficacy of the treatment.

Consequently, in one embodiment, the compound or composition providedherein is administered once a week in a single or divided doses in athree to six week cycle with a rest period of about 1 to about 30 days.In another embodiment, the compound or composition provided herein isadministered once a week in a single or divided doses for one week, twoweeks, three weeks, four weeks, five weeks or six weeks with a restperiod of 1, 3, 5, 7, 9, 12, 14, 16, 18, 20, 21, 22, 24, 26, 28, 29 or30 days. In some embodiments, the waiting period is 14 days. In someembodiments, the waiting period is 28 days. The frequency, number andlength of dosing cycles can be increased or decreased. Thus, anotherembodiment encompasses the administration of the compound or compositionprovided herein for more cycles than are typical when the compound orcomposition is administered alone.

In one embodiment, the method provided herein comprises: i)administering a dose of about 1 mg/m²-150 mg/m² of the compound orcomposition provided herein to a patient; ii) waiting a period of atleast one day where the mammal is not administered the compound orcomposition; and iii) administering another dose of about 1 mg/m²-150mg/m² of the compound or composition provided herein to the patient. Inone embodiment, steps ii)-iii) are repeated a plurality of times.

In one embodiment, the method provided herein comprises: i)administering a dose of about 1 mg/m²-75 mg/m² of the compound orcomposition provided herein to a patient; ii) waiting a period of atleast one day where the mammal is not administered the compound orcomposition; and iii) administering another dose of about 1 mg/m²-75mg/m² of the compound or composition provided herein to the patient. Inone embodiment, steps ii)-iii) are repeated a plurality of times.

In one embodiment, the method provided herein comprises: i)administering a dose of about 1 mg/m²-75 mg/m² of the compound orcomposition provided herein to a patient; ii) waiting a period of atleast two days where the mammal is not administered the compound orcomposition; and iii) administering another dose of about 1 mg/m²-75mg/m² of the compound or composition provided herein to the patient. Inone embodiment, steps ii)-iii) are repeated a plurality of times. In yetanother embodiment, the method comprises administering a dose of about 1mg/m²-50 mg/m², 1 mg/m²-60 mg/m², 10 mg/m²-60 mg/m² or about 1 mg/m²-48mg/m² of the compound or composition provided herein in steps i) andiii). In yet another embodiment, the method comprises administering adose of about 15 mg/m², 48 mg/m², 50 mg/m², 60 mg/m² or 75 mg/m² of thecompound or composition provided herein in steps i) and iii).

In one embodiment, the method provided herein comprises: i)administering a dose of about 1 mg/m²-75 mg/m² of the compound orcomposition provided herein to a patient; ii) waiting a period of atleast 7 days where the mammal is not administered the compound orcomposition; and iii) administering another dose of about 1 mg/m²-75mg/m² of the compound or composition provided herein to the patient. Inone embodiment, steps ii)-iii) are repeated a plurality of times. In yetanother embodiment, the method comprises administering a dose of about 1mg/m²-50 mg/m² or about 1 mg/m²48 mg/m² of the compound or compositionprovided herein in steps i) and iii). In yet another embodiment, themethod comprises administering a dose of about 15 mg/m², 48 mg/m², 50mg/m², 60 mg/m² or 75 mg/m² of the compound or composition providedherein in steps i) and iii).

In one embodiment, the method provided herein comprises: i)administering a dose of about 1 mg/m²-75 mg/m² of the compound orcomposition provided herein to a patient; ii) waiting a period of atleast 14 days where the mammal is not administered the compound orcomposition; and iii) administering another dose of about 1 mg/m²-75mg/m² of the compound or composition provided herein to the patient. Inone embodiment, steps ii)-iii) are repeated a plurality of times.

In one embodiment, the method provided herein comprises: i)administering a dose of about 10 mg/m²-60 mg/m² of the compound orcomposition provided herein to a patient; ii) waiting a period of atleast one day where the mammal is not administered the compound orcomposition; and iii) administering another dose of about 10 mg/m²-60mg/m² of the compound or composition provided herein to the patient. Inone embodiment, steps ii)-iii) are repeated a plurality of times.

In one embodiment, the method provided herein comprises: i)administering a dose of about 10 mg/m²-60 mg/m² of the compound orcomposition provided herein to a patient; ii) waiting a period of atleast 7 days where the mammal is not administered the compound orcomposition; and iii) administering another dose of about 10 mg/m²-60mg/m² of the compound or composition provided herein to the patient. Inone embodiment, steps ii)-iii) are repeated a plurality of times.

In one embodiment, the method provided herein comprises: i)administering a dose of about 10 mg/m²-60 mg/m² of the compound orcomposition provided herein to a patient; ii) waiting a period of atleast 14 days where the mammal is not administered the compound orcomposition; and iii) administering another dose of about 10 mg/m²-60mg/m² of the compound or composition provided herein to the patient. Inone embodiment, steps ii)-iii) are repeated a plurality of times.

In one embodiment, the method provided herein comprises: i)administering a dose of about 1 mg/m²-48 mg/m² of the compound orcomposition provided herein to a patient; ii) waiting a period of atleast one day where the mammal is not administered compound orcomposition; and iii) administering another dose of about 1 mg/m²-48mg/m² of the compound or composition to the patient. In one embodiment,steps ii)-iii) are repeated a plurality of times.

In one embodiment, the method provided herein comprises: i)administering a dose of about 1 mg/m²-24 mg/m² of the compound orcomposition provided herein to a patient; ii) waiting a period of atleast one day where the mammal is not administered the compound orcomposition; and iii) administering another dose of about 1 mg/m²-24mg/m² of the compound or composition to the patient. In one embodiment,steps ii)-iii) are repeated a plurality of times.

In another embodiment, the method comprises administering a dose ofabout 3 mg/m²-24 mg/m² of the compound or composition in steps i) andiii). In yet another embodiment, the method comprises administering adose of about 15 mg/m² in steps i) and iii). In yet another embodiment,the method comprises administering a dose of about 1 mg/m²-40 mg/m²,about 1.5 mg/m²-30 mg/m², about 2 mg/m²-25 mg/m² or about 3 mg/m²-24mg/m² of the compound or composition provided herein in steps i) andiii).

In one embodiment, the method provided herein comprises: i)administering a dose of about 1 mg/m²-60 mg/m² or about 10 mg/m²-60mg/m² of the compound or composition provided herein to a patient; ii)waiting a period of at least one day where the mammal is notadministered the compound or composition; and iii) administering anotherdose of about 1 mg/m²-60 mg/m² or about 10 mg/m²-60 mg/m² of thecompound or composition provided herein to the patient. In oneembodiment, steps ii)-iii) are repeated a plurality of times. In oneembodiment, the waiting period is 6 days. In one embodiment, stepsii)-iii) are repeated a plurality of times. In one embodiment, thewaiting period is 7 days. In one embodiment, steps ii)-iii) are repeateda plurality of times. In one embodiment, the waiting period is 14 days.In one embodiment, the waiting period is 18 days. In one embodiment, thewaiting period is 21 days.

In one embodiment, the method provided herein comprises i) administeringa dose of about 1 mg/m²-75 mg/m² of the compound or composition providedherein to a patient once every week for three weeks and ii) waiting fora period of 14 days. In one embodiment, the method provided hereincomprises i) administering a dose of about 1 mg/m²-60 mg/m² or about 10mg/m²-60 mg/m² of the compound or composition provided herein to apatient once every week for three weeks and ii) waiting for a period of14 days.

In one embodiment, steps i)-ii) are repeated a plurality of times.

In one embodiment, the method provided herein comprises i) administeringa dose of about 10 mg/m²-48 mg/m² of the compound or compositionprovided herein to a patient once every week for three weeks and ii)waiting for a period of 14 days. In one embodiment, steps ii)-ii) arerepeated a plurality of times.

In one embodiment, the method provided herein comprises i) administeringa dose of about 15 mg/m² of the compound or composition provided hereinto a patient once every week for three weeks, ii) waiting for a periodof 7 days and iii) administering another dose of about 15 mg/m² of thecompound or composition provided herein to a patient once every week forthree weeks. In one embodiment, steps ii)-iii) are repeated a pluralityof times.

In one embodiment, the method provided herein comprises i) administeringa dose of about 15 mg/m² of the compound or composition provided hereinto a patient once every week for three weeks, ii) waiting for a periodof 14 days and iii) administering another dose of about 15 mg/m² of thecompound or composition provided herein to a patient once every week forthree weeks. In one embodiment, steps ii)-iii) are repeated a pluralityof times.

In one embodiment, the method provided herein comprises administering adose of about 60 mg/m² of the compound or composition provided herein toa patient once every three weeks. In one embodiment, the method providedherein comprises administering a dose of about 48 mg/m² of the compoundor composition provided herein to a patient once every three weeks. Inone embodiment, the method provided herein is for treatment of solidtumors. In one embodiment, the method is for treatment ofleiomyosarcoma, melanoma, mesothelioma, mesothelioma, nasopharyngeal,renal cell cancer, salivary gland cancer, lung cancer, adenocarcinoma(unknown origin), ovarian cancer, colon cancer, neuroendocrine, ormüllerian tumor.

In another embodiment, the method provided herein comprises: i)administering a dose of about 3 mg/m²-24 mg/m² of the compound orcomposition to a mammal once a week for 3 weeks (e.g. dosing in days 1,8 and 15); ii) waiting a period of at least 28 days where the mammal isnot administered the compound or composition; and iii) administeringanother dose of about 3 mg/m²-24 mg/m² of the compound or composition tothe mammal once a week for 3 weeks. In one embodiment, steps ii)-iii)are repeated a plurality of times.

In another embodiment, the method provided herein comprises: i)administering a dose of about 10 mg/m²-60 mg/m² of the compound orcomposition to a mammal once a week for 3 weeks (e.g. dosing in days 1,8 and 15); ii) waiting a period of at least 28 days where the mammal isnot administered the compound or composition; and iii) administeringanother dose of about 10 mg/m²-60 mg/m² of the compound or compositionto the mammal once a week for 3 weeks. In one embodiment, steps ii)-iii)are repeated a plurality of times.

In another embodiment, the method provided herein comprises: i)administering a dose of about 3 mg/m²-24 mg/m² of the compound orcomposition to a mammal once a week for 3 weeks (e.g. dosing in days 1,8 and 15); ii) waiting a period of 28 days where the mammal is notadministered the compound or composition; and iii) administering anotherdose of about 3 mg/m²-24 mg/m² of the compound or composition to themammal once a week for 3 weeks. In one embodiment, steps ii)-iii) arerepeated a plurality of times.

In another embodiment, the method provided herein comprises: i)administering a dose of about 10 mg/m²-60 mg/m² of the compound orcomposition to a mammal once a week for 3 weeks (e.g. dosing in days 1,8 and 15); ii) waiting a period of 28 days where the mammal is notadministered the compound or composition; and iii) administering anotherdose of about 10 mg/m²-60 mg/m² of the compound or composition to themammal once a week for 3 weeks. In one embodiment, steps ii)-iii) arerepeated a plurality of times.

In another embodiment, the method provided herein comprises: i)administering a dose of about 1 mg/m²-75 mg/m² of the compound orcomposition to a mammal twice a week for 2 weeks (dosing in days 1, 4, 8and 1); ii) waiting a period of at least 7 days where the mammal is notadministered the compound or composition; and iii) administering anotherdose of about 1 mg/m²-75 mg/m² of the compound or composition to themammal twice a week for 2 weeks (dosing in days 1, 4, 8 and 11). In oneembodiment, steps ii)-iii) are repeated a plurality of times.

In another embodiment, the method provided herein comprises: i)administering a dose of about 1 mg/m²-75 mg/m² of the compound orcomposition to a mammal twice a week for 2 weeks (dosing in days 1, 4, 8and 11); ii) waiting a period of at least 14 days where the mammal isnot administered the compound or composition; and iii) administeringanother dose of about 1 mg/m²-75 mg/m² of the compound or composition tothe mammal twice a week for 2 weeks (dosing in days 1, 4, 8 and 11). Inone embodiment, steps ii)-iii) are repeated a plurality of times.

In another embodiment, the method provided herein comprises: i)administering a dose of about 3 mg/m²-24 mg/m² of the compound orcomposition to a mammal twice a week for 2 weeks (dosing in days 1, 4, 8and 11); ii) waiting a period of at least 28 days where the mammal isnot administered the compound or composition; and iii) administeringanother dose of about 3 mg/m²-24 mg/m² of the compound or composition tothe mammal twice a week for 2 weeks (dosing in days 1, 4, 8 and 11). Inone embodiment, steps ii)-iii) are repeated a plurality of times.

In another embodiment, the method provided herein comprises: i)administering a dose of about 1 mg/m²-75 mg/m² of the compound orcomposition to a mammal twice a week for 2 weeks (dosing in days 1, 4, 8and 11); ii) waiting a period of at least 28 days where the mammal isnot administered the compound or composition; and iii) administeringanother dose of about 1 mg/m²-75 mg/m² of the compound or composition tothe mammal twice a week for 2 weeks (dosing in days 1, 4, 8 and 11). Inone embodiment, steps ii)-iii) are repeated a plurality of times.

In another embodiment, the method provided herein comprises: i)administering a dose of about 1 mg/m²-75 mg/m² of the compound orcomposition to a mammal twice a week for 2 weeks (dosing in days 1, 4, 8and 11); ii) waiting a period of at least 14 days where the mammal isnot administered the compound or composition; and iii) administeringanother dose of about 1 mg/m²-75 mg/m² of the compound or composition tothe mammal twice a week for 2 weeks (dosing in days 1, 4, 8 and 11). Inone embodiment, steps ii)-iii) are repeated a plurality of times.

In another embodiment, the method provided herein comprises: i)administering a dose of about 10 mg/m²-60 mg/m² of the compound orcomposition to a mammal twice a week for 2 weeks (dosing in days 1, 4, 8and 11); ii) waiting a period of at least 28 days where the mammal isnot administered the compound or composition; and iii) administeringanother dose of about 10 mg/m²-60 mg/m² of the compound or compositionto the mammal twice a week for 2 weeks (dosing in days 1, 4, 8 and 11).In one embodiment, steps ii)-iii) are repeated a plurality of times.

In another embodiment, the method provided herein comprises: i)administering a dose of about 3 mg/m²-24 mg/m² of the compound orcomposition to a mammal twice a week for 2 weeks (dosing in days 1, 4, 8and 11); ii) waiting a period of 28 days where the mammal is notadministered the compound or composition; and iii) administering anotherdose of about 3 mg/m²-24 mg/m² of the compound or composition to themammal twice a week for 2 weeks (dosing in days 1, 4, 8 and 11). In oneembodiment, steps ii)-iii) are repeated a plurality of times.

In another embodiment, the method provided herein comprises: i)administering a dose of about 10 mg/m²-60 mg/m² of the compound orcomposition to a mammal twice a week for 2 weeks (dosing in days 1, 4, 8and 11); ii) waiting a period of 28 days where the mammal is notadministered the compound or composition; and iii) administering anotherdose of about 10 mg/m²-60 mg/m² of the compound or composition to themammal twice a week for 2 weeks (dosing in days 1, 4, 8 and 11). In oneembodiment, steps ii)-iii) are repeated a plurality of times.

In one embodiment, the method for treatment of solid tumors providedherein comprises i) administering a dose of about 15 mg/m² of thecompound or composition provided herein to a patient once every week forthree weeks and ii) waiting for a period of 14 days. In one embodiment,steps ii)-iii) are repeated a plurality of times.

In one embodiment, the method for treatment of solid tumors providedherein comprises i) administering a dose of about 15 mg/m² of thecompound or composition provided herein to a patient once every week forthree weeks and ii) waiting for a period of 7 days. In one embodiment,steps ii)-iii) are repeated a plurality of times.

In another embodiment, provided herein is a method for treatment ofsolid tumors comprising administering a dose of about 48 mg/m², 60 mg/m²or 75 mg/m² of the compound or composition provided herein to a patientonce every three weeks.

In another embodiment, provided herein is a method for treatment ofleukemia comprising administering a dose of about 50 mg/m², 60 mg/m² or72 mg/m² of the compound or composition provided herein to a patientonce every three weeks.

In another embodiment, provided herein is a method for treatment ofleukemia comprising administering a dose of about 40 mg/m² or 50 mg/m²of the compound or composition provided herein to a patient once everythree weeks.

In one embodiment, leukemia is acute leukemia. In one embodiment,leukemia is relapsed and/or refractory acute leukemia. In oneembodiment, leukemia is relapsed acute leukemia. In one embodiment,leukemia is refractory acute leukemia.

In another embodiment, the dosing method comprises administering a doseof the compound or composition twice a week (dosing on days 1, 4, 8 and11) to a mammal.

In another embodiment, the dosing method comprises administering a oncea week dose of the compound or composition to a mammal. In anotherembodiment, the dosing method comprises administering a dose of thecompound or composition to a mammal once every two weeks. In anotherembodiment, the dosing method comprises administering a dose of thecompound or composition to a mammal once every three weeks. In anotherembodiment, the dosing method comprises administering a dose of thecompound or composition to a mammal once every four weeks.

In another embodiment, the dosing method comprises a cycle wherein thecycle comprises administering a dose of the compound or composition to amammal once a week for three weeks followed by a period of at least 14days where no compound or composition is administered to the mammal andwherein the cycle is repeated a plurality of times. In anotherembodiment, the period where no compound or composition is administeredis 18 days. In another embodiment, the period where no compound orcomposition is administered is 21 days. In another embodiment, theperiod where no compound or composition is administered is 28 days.

In the above methods, if the waiting period were 6 days, then theinitial dose of the compound or composition is administered on Day 1(step i); the waiting period is six days (step ii); and the followingdose of the compound or composition is administered on Day 8 (step iii).Other exemplary time periods include 2 days, 3 days, 5 days, 7 days, 10days, 12 days, 13 days, 14 days, 15 days, 17 days, 20 days, 21 days and28 days. In one embodiment, the waiting period is until there issufficient bone marrow recovery. In another embodiment, the waitingperiod is at least 2 days and steps ii) through iii) are repeated atleast three times. In another embodiment, the waiting period is at least2 days and steps ii) through iii) are repeated at least five times. Inanother embodiment, the waiting period is at least 3 days and steps ii)through iii) are repeated at least three times. In another embodiment,the waiting period is at least 3 days and steps ii) through iii) arerepeated at least five times. In another embodiment, the waiting periodis at least 6 days and steps ii) through iii) are repeated at leastthree times. In another embodiment, the waiting period is at least 6days and steps ii) through iii) are repeated at least five times. Inanother embodiment, the waiting period is at least 14 days and steps ii)through iii) are repeated at least three times. In another embodiment,the waiting period is at least 14 days and steps ii) through iii) arerepeated at least five times. In another embodiment, the waiting periodis at least 20 days and steps ii) through iii) are repeated at leastthree times. In another embodiment, the waiting period is at least 20days and steps ii) through iii) are repeated at least five times. Inanother embodiment, the waiting period is at least 28 days and steps ii)through iii) are repeated at least three times. In another embodiment,the waiting period is at least 28 days and steps ii) through iii) arerepeated at least five times.

In another embodiment, the method comprises administering a dose of 1mg/m²-75 mg/m² of the compound or composition to a patient once a weekwherein the one-week period comprises a treatment cycle and thetreatment cycle is repeated at least three times. In another embodiment,the method comprises administering a dose of 1 mg/m²-60 mg/m² of thecompound or composition to a patient once a week wherein the one-weekperiod comprises a treatment cycle and the treatment cycle is repeatedat least three times. In another embodiment, the method comprisesadministering a dose of 1 mg/m²-48 mg/m² of the compound or compositionto a patient once a week wherein the one-week period comprises atreatment cycle and the treatment cycle is repeated at least threetimes. In another embodiment, the method comprises administering a doseof 1 mg/m²-24 mg/m² of the compound or composition to a patient once aweek wherein the one-week period comprises a treatment cycle and thetreatment cycle is repeated at least three times. In another embodiment,the dose is about 2 mg/m²-40 mg/m² of the compound or composition once aweek wherein the one-week period comprises a treatment cycle and thetreatment cycle is repeated at least three times. In another embodiment,the dose is about 3 mg/m²-24 mg/m² of the compound or composition once aweek wherein the one-week period comprises a treatment cycle and thetreatment cycle is repeated at least three times. In another embodiment,the dose is about 15 mg/m² of the compound or composition once a weekwherein the one-week period comprises a treatment cycle and thetreatment cycle is repeated at least three times.

In some embodiments, the method comprises administering a dose of about10 mg/m²-60 mg/m² of the compound or composition to a patient once aweek (e.g. dosing in days 1, 8 and 15) wherein the one-week periodcomprises a treatment cycle and the treatment cycle is repeated at leastthree times followed by a waiting period of at least 28 days. In someembodiments, the method comprises administering a dose of about 10mg/m²-48 mg/m² of the compound or composition to a patient once a week(e.g. dosing in days 1, 8 and 15) wherein the one-week period comprisesa treatment cycle and the treatment cycle is repeated at least threetimes followed by a waiting period of at least 28 days. In someembodiments, the method comprises administering a dose of about 10mg/m²-48 mg/m² of the compound or composition to a patient twice a week(dosing in days 1, 4, 8, and 11) wherein the one-week period comprises atreatment cycle and the treatment cycle is repeated at least three timesfollowed by a waiting period of at least 28 days. In some embodiments,the method comprises administering a dose of about 10 mg/m²-60 mg/m² ofthe compound or composition to a patient once a week (e.g. dosing indays 1, 8 and 15) wherein the one-week period comprises a treatmentcycle and the treatment cycle is repeated at least three times followedby a waiting period of 28 days. In some embodiments, the methodcomprises administering a dose of about 10 mg/m²-48 mg/m² of thecompound or composition to a patient once a week (e.g. dosing in days 1,8 and 15) wherein the one-week period comprises a treatment cycle andthe treatment cycle is repeated at least three times followed by awaiting period of 28 days. In some embodiments, the method comprisesadministering a dose of about 10 mg/m²-48 mg/m² of the compound orcomposition to a patient twice a week (dosing in days 1, 4, 8, and 11)wherein the one-week period comprises a treatment cycle and thetreatment cycle is repeated at least three times followed by a waitingperiod of 28 days. In some embodiments, the method comprisesadministering a dose of about 3 mg/m²-24 mg/m² of the compound orcomposition to a patient once a week (e.g. dosing in days 1, 8 and 15)wherein the one-week period comprises a treatment cycle and thetreatment cycle is repeated at least three times followed by a waitingperiod of at least 28 days. In some embodiments, the method comprisesadministering a dose of about 3 mg/m²-24 mg/m² of the compound orcomposition to a patient twice a week (dosing in days 1, 4, 8, and 11)wherein the one-week period comprises a treatment cycle and thetreatment cycle is repeated at least three times followed by a waitingperiod of at least 28 days. In some embodiments, the method comprisesadministering a dose of about 3 mg/m²-24 mg/m² of the compound orcomposition to a patient once a week (e.g. dosing in days 1, 8 and 15)wherein the one-week period comprises a treatment cycle and thetreatment cycle is repeated at least three times followed by a waitingperiod of 28 days. In some embodiments, the method comprisesadministering a dose of about 3 mg/m²-24 mg/m² of the compound orcomposition to a patient twice a week (dosing in days 1, 4, 8, and 11)wherein the one-week period comprises a treatment cycle and thetreatment cycle is repeated at least three times followed by a waitingperiod of 28 days. All methods and dosages described herein apply to thetreatment or prevention of cancer.

In certain embodiments, the methods provided herein comprise monitoringan amount of N-desmethyl-SNS-595 in a subject administered with apharmaceutical composition comprising SNS-595. The subject can be anysubject for which such monitoring might be useful. For example, anexemplary method for detection of N-desmethyl SNS-595 generated in vitrois described in the Examples section. Other methods known to one ofskill in the art could be used.

5.5 Second Active Agents

In the methods and pharmaceutical compositions provided herein, thecompound or composition provided herein can be used with or combinedwith other pharmacologically active compounds (“second active agents”).It is believed that certain combinations work synergistically in thetreatment of particular types of cancers. The compound or compositionprovided herein can also work to alleviate adverse effects associatedwith certain second active agents, and some second active agents can beused to alleviate adverse effects associated with the compound orcomposition provided herein.

One or more second active ingredients or agents can be used in themethods and pharmaceutical compositions provided herein together withthe compound or composition provided herein. Second active agents can belarge molecules (e.g., proteins) or small molecules (e.g., syntheticinorganic, organometallic or organic molecules).

Examples of large molecule active agents include, but are not limitedto, hematopoietic growth factors, cytokines, and monoclonal andpolyclonal antibodies, particularly therapeutic antibodies to cancerantigens. Typical large molecule active agents are biological molecules,such as naturally occurring or artificially made proteins. Proteins thatare particularly useful in the methods and compositions provided hereininclude proteins that stimulate the survival and/or proliferation ofhematopoietic precursor cells and immunologically active poietic cellsin vitro or in vivo. Others stimulate the division and differentiationof committed erythroid progenitors in cells in vitro or in vivo.Particular proteins include, but are not limited to: interleukins, suchas IL-2 (including recombinant IL-II (“rIL2”) and canarypox IL-2),IL-10, IL-12, and IL-18; interferons, such as interferon alfa-2a,interferon alfa-2b, interferon alfa-n1, interferon alfa-n3, interferonbeta-I a, and interferon gamma-I b; G-CSF (Granulocytecolony-stimulating factor) and GM-CSF (Granulocyte-macrophagecolony-stimulating factor); and EPO (Erythropoietin).

Particular proteins that can be used in the methods and compositionsinclude, but are not limited to: filgrastim, which is sold in the UnitedStates under the trade name Neupogen® (Amgen, Thousand Oaks, Calif.);sargramostim, which is sold in the United States under the trade nameLeukine® (Immunex, Seattle, Wash.); and recombinant EPO, which is soldin the United States under the trade name Epogen® (Amgen, Thousand Oaks,Calif.).

Recombinant and mutated forms of GM-CSF can be prepared as described inU.S. Pat. Nos. 5,391,485; 5,393,870; and 5,229,496; all of which areincorporated herein by reference. Recombinant and mutated forms of G-CSFcan be prepared as described in U.S. Pat. Nos. 4,810,643; 4,999,291;5,528,823; and 5,580,755; all of which are incorporated herein byreference.

Also provided for use in combination with the compounds and compositionsprovided herein are native, naturally occurring, and recombinantproteins. Further encompassed are mutants and derivatives (e.g.,modified forms) of naturally occurring proteins that exhibit, in vivo,at least some of the pharmacological activity of the proteins upon whichthey are based. Examples of mutants include, but are not limited to,proteins that have one or more amino acid residues that differ from thecorresponding residues in the naturally occurring forms of the proteins.Also encompassed by the term “mutants” are proteins that lackcarbohydrate moieties normally present in their naturally occurringforms (e.g., nonglycosylated forms). Examples of derivatives include,but are not limited to, pegylated derivatives and fusion proteins, suchas proteins formed by fusing IgG1 or IgG3 to the protein or activeportion of the protein of interest. See, e.g., Penichet, M. L. andMorrison, S. L., J. Immunol. Methods 248:91-101 (2001). Antibodies thatcan be used in combination with SNS-595 include monoclonal andpolyclonal antibodies. Examples of antibodies include, but are notlimited to, trastuzumab (Herceptin®), rituximab (Rituxan®), bevacizumab(Avastin™), pertuzumab (Omnitarg™), tositumomab (Bexxar®), edrecolomab(Panorex®), and G250. SNS-595 can also be combined with or used incombination with, anti-TNF-α antibodies.

Large molecule active agents may be administered in the form ofanti-cancer vaccines. For example, vaccines that secrete or cause thesecretion of, cytokines such as IL-2, G-CSF, and GM-CSF can be used inthe methods and pharmaceutical compositions provided. See, e.g., Emens,L. A., et al., Curr. Opinion Mol. Ther. 3(1):77-84 (2001).

Second active agents that are small molecules can also be used toalleviate adverse effects associated with the administration of thecompound or composition provided herein. However, like some largemolecules, many are believed to be capable of providing a synergisticeffect when administered with (e.g., before, after or simultaneously)the compound or composition provided herein. Examples of small moleculesecond active agents include, but are not limited to, anti-canceragents, antibiotics, immunosuppressive agents, and steroids.

Examples of anti-cancer agents include, but are not limited to:acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin;aldesleukin; altretamine; ambomycin; ametantrone acetate; amsacrine;anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa;azotomycin; batimastat; benzodepa; bicalutamide; bisantrenehydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate;brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone;caracemide; carbetimer; carboplatin; carmustine; carubicinhydrochloride; carzelesin; cedefingol; celecoxib (COX-2 inhibitor);chlorambucil; cirolemycin; cisplatin; cladribine; crisnatol mesylate;cyclophosphamide; cytarabine; dacarbazine; dactinomycin; daunorubicinhydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguaninemesylate; diaziquone; docetaxel; doxorubicin; doxorubicin hydrochloride;droloxifene; droloxifene citrate; dromostanolone propionate; duazomycin;edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin;enpromate; epipropidine; epirubicin hydrochloride; erbulozole;esorubicin hydrochloride; estramustine; estramustine phosphate sodium;etanidazole; etoposide; etoposide phosphate; etoprine; fadrozolehydrochloride; fazarabine; fenretinide; floxuridine; fludarabinephosphate; fluorouracil; fluorocitabine; fosquidone; fostriecin sodium;gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicinhydrochloride; ifosfamide; ilmofosine; iproplatin; irinotecan;irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolideacetate; liarozole hydrochloride; lometrexol sodium; lomustine;losoxantrone hydrochloride; masoprocol; maytansine; mechlorethaminehydrochloride; megestrol acetate; melengestrol acetate; melphalan;menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine;meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin;mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolicacid; nocodazole; nogalamycin; ormaplatin; oxisuran; paclitaxel;pegaspargase; peliomycin; pentamustine; peplomycin sulfate;perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine;procarbazine hydrochloride; puromycin; puromycin hydrochloride;pyrazofurin; riboprine; safingol; safingol hydrochloride; semustine;simtrazene; sparfosate sodium; sparsomycin; spirogermaniumhydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin;sulofenur; talisomycin; tecogalan sodium; taxotere; tegafur;teloxantrone hydrochloride; temoporfin; teniposide; teroxirone;testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;tirapazamine; toremifene citrate; trestolone acetate; triciribinephosphate; trimetrexate; trimetrexate glucuronate; triptorelin;tubulozole hydrochloride; uracil mustard; uredepa; vapreotide;verteporfin; vinblastine sulfate; vincristine sulfate; vindesine;vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; and zorubicinhydrochloride.

Other anti-cancer drugs include, but are not limited to: 20-epi-1, 25dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin;acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists;altretamine; ambamustine; amidox; amifostine; aminolevulinic acid;amrubicin; amsacrine; anagrelide; anastrozole; andrographolide;angiogenesis inhibitors; antagonist D; antagonist G; antarelix;anti-dorsalizing morphogenetic protein-1; antiandrogen, prostaticcarcinoma; antiestrogen; antineoplaston; antisense oligonucleotides;aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators;apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine;atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3;azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol;batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine;beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid;bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine;bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane;buthionine sulfoximine; calcipotriol; calphostin C; camptothecinderivatives; capecitabine; carboxamide-amino-triazole;carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor;carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropinB; cetrorelix; chlorlns; chloroquinoxaline sulfonamide; cicaprost;cis-porphyrin; cladribine; clomifene analogues; clotrimazole;collismycin A; collismycin B; combretastatin A4; combretastatinanalogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8;cryptophycin A derivatives; curacin A; cyclopentanthraquinones;cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor;cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin;dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone;didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine;dihydrotaxol, 9-; dioxamycin; diphenyl spiromustine; docetaxel;docosanol; dolasetron; doxifluridine; doxorubicin; droloxifene;dronabinol; duocarmycin SA; ebselen; ecomustine; edelfosine;edrecolomab; eflomithine; elemene; emitefur; epirubicin; epristeride;estramustine analogue; estrogen agonists; estrogen antagonists;etanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine;fenretinide; filgrastim; finasteride; flavopiridol; flezelastine;fluasterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex;formestane; fostriecin; fotemustine; gadolinium texaphyrin; galliumnitrate; galocitabine; ganirelix; gelatinase inhibitors; gemcitabine;glutathione inhibitors; hepsulfam; heregulin; hexamethylenebisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene;idramantone; ilmofosine; ilomastat; imatinib (e.g., Gleevec®);imiquimod; immunostimulant peptides; insulin-like growth factor-1receptor inhibitor; interferon agonists; interferons; interleukins;iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact; irsogladine;isobengazole; isohomohalicondrin B; itasetron; jasplakinolide;kahalalide F; lamellarin-N triacetate; lanreotide; leinamycin;lenograstim; lentinan sulfate; leptolstatin; letrozole; leukemiainhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; loxoribine; lurtotecan; lutetiumtexaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A;marimastat; masoprocol; maspin; matrilysin inhibitors; matrixmetalloproteinase inhibitors; menogaril; merbarone; meterelin;methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine;mirimostim; mitoguazone; mitolactol; mitomycin analogues; mitonafide;mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofarotene;molgramostim; Erbitux, human chorionic gonadotrophin; monophosphoryllipid A+myobacterium cell wall sk; mopidamol; mustard anticancer agent;mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; nilutamide; nisamycin; nitric oxidemodulators; nitroxide antioxidant; nitrullyn; oblimersen (Genasense®);O⁶-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues;paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid;panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase;peldesine; pentosan polysulfate sodium; pentostatin; pentrozole;perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rohitukine;romurtide; roquinimex; rubiginone B1; ruboxyl; safingol; saintopin;SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics; semustine;senescence derived inhibitor 1; sense oligonucleotides; signaltransduction inhibitors; sizofuran; sobuzoxane; sodium borocaptate;sodium phenylacetate; solverol; somatomedin binding protein; sonermin;sparfosic acid; spicamycin D; spiromustine; splenopentin; spongistatin1; squalamine; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; tallimustine; tamoxifen methiodide; tauromustine;tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomeraseinhibitors; temoporfin; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; translation inhibitors; tretinoin;triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron;turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors;ubenimex; urogenital sinus-derived growth inhibitory factor; urokinasereceptor antagonists; vapreotide; variolin B; velaresol; veramine;verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole;zanoterone; zeniplatin; zilascorb; and zinostatin stimalamer.

Specific second active agents include, but are not limited to,rituximab; oblimersen (Genasense®); remicade; docetaxel; celecoxib;melphalan; dexamethasone (Decadron®); steroids; gemcitabine;cisplatinum; temozolomide; etoposide; cyclophosphamide; temodar;carboplatin; procarbazine; gliadel; tamoxifen; topotecan; methotrexate;Arisa®; taxol; taxotere; fluorouracil; leucovorin; irinotecan; xeloda;CPT-11; interferon alpha; pegylated interferon alpha (e.g., PEGINTRON-A); capecitabine; cisplatin; thiotepa; fludarabine; carboplatin;liposomal daunorubicin; cytarabine; doxetaxol; pacilitaxel; vinblastine;IL-2; GM-CSF; dacarbazine; vinorelbine; zoledronic acid; palmitronate;biaxin; busulphan; prednisone; bisphosphonate; arsenic trioxide;vincristine; doxorubicin (Doxil®); paclitaxel; ganciclovir; adriamycin;estramustine sodium phosphate (Emcyt®); sulindac; and etoposide.

In certain embodiments, the second active agent is etoposide;daunomycin; actinomycin D; mitomycin C; cisplatin; carboplatin;premetrexed; methotrexate; Ara-C; 5-Fu; wortmannin; gemcitabin;geldanamycin or a combination thereof.

5.6 Combination Therapy with a Second Active Agent

In certain embodiments, the method provided herein comprisesadministering a compound or composition provided herein in combinationwith one or more second active agents, and/or in combination withradiation therapy or surgery. The administration of the compound orcomposition and the second active agents to a patient can occursimultaneously or sequentially by the same or different routes ofadministration. The suitability of a particular route of administrationemployed for a particular active agent will depend on the active agentitself (e.g., whether it can be administered orally without decomposingprior to entering the blood stream) and the disease being treated.Recommended routes of administration for the second active agents areknown to those of ordinary skill in the art. See, e.g., Physicians' DeskReference, 1755-1760 (56^(th) ed., 2002).

In one embodiment, the second active agent is administered intravenouslyor subcutaneously and once or twice daily in an amount of from about 1to about 1,000 mg, from about 5 to about 500 mg, from about 10 to about375 mg or from about 50 to about 200 mg. In one embodiment, the secondactive agent is rituximab, oblimersen (Genasense®), GM-CSF, G-CSF, EPO,taxotere, irinotecan, dacarbazine, transretinoic acid, topotecan,pentoxifylline, ciprofloxacin, dexamethasone, vincristine, doxorubicin,COX-2 inhibitor, IL2, IL8, IL18, IFN, Ara-C, vinorelbine or acombination thereof. In certain embodiments, the second active agent isetoposide, daunomycin, actinomycin D, mitomycin C, cisplatin,carboplatin, premetrexed, methotrexate, Ara-C (cytarabine), 5-FU(Fluorouracil), wortnannin, geldanamycin, gemcitabine or a combinationthereof.

In another embodiment, provided herein are methods of treating,preventing and/or managing hematologic malignancies, which compriseadministering a compound or composition provided herein in conjunctionwith (e.g., before, during or after) conventional therapy including, butnot limited to, surgery, immunotherapy, biological therapy, radiationtherapy or other non-drug based therapy presently used to treat, preventor manage cancer.

In certain embodiments, the second active agent is co-administered witha compound or composition provided herein or administered with 1-50hours delay. In certain embodiments, a compound or composition providedherein is administered first followed by administration with the secondactive agent with 1-50 hours delay. In other embodiments, the secondactive agent is administered first followed by administration of acompound or composition provided herein with 1-50 hours delay. In someembodiment, the delay is 24 hours.

In one embodiment, a compound or composition provided herein can beadministered in an amount of from about 1 to about 150 mg/m², 1 to about75 mg/m², 1 to about 60 mg/m², 1 to about 48 mg/m², 1 to about 24 mg/m²,1 to about 50 mg/m², about 1 to about 40 mg/m², about 1 to about 30mg/m², about 3 to about 30 mg/m², about 3 to about 24 mg/m² alone or incombination with a second active agent disclosed herein (see, e.g.,section 5.6), prior to, during, or after the use of conventionaltherapy.

In another embodiment, the method provided herein comprises: a)administering to a patient in need thereof, a dose of about 1 mg/m²-75mg/m² of a compound or composition provided herein and b) administeringa therapeutically effective amount of a supportive care agent.

In another embodiment, the method provided herein comprises: a)administering to a patient in need thereof, a dose of about 10 mg/m²-60mg/m² of a compound or composition provided herein and b) administeringa therapeutically effective amount of a supportive care agent.

The supportive care agent is any substance that treats, prevents,manages, avoids or reduces an adverse or unwanted effect from treatmentwith a compound or composition provided herein and is administeredaccording to the appropriate dosing regimen for that substance. Forexample, different supportive care agents for treating nausea havedifferent dosing regimens. While some are administered prophylactically,others are co-administered with a compound or composition providedherein while still others are administered after the administration ofthe compound or composition. Illustrative examples of supportive careagents their doses and dosing regimens are found in The Physician's DeskReference. Some exemplary support care agents are disclosed in U.S.Application Pub. No. 2006-0025437, which incorporated by reference inits entirety.

5.7 Pharmaceutical Compositions and Dosage Forms

The methods provided herein use pharmaceutical compositions containingthe compound or compositions provided herein and pharmaceuticallyacceptable carriers, such as diluents or adjuvants, or in combinationwith other active ingredients, such as another anti-cancer agent. Inclinical practice, the compound or composition may be administered byany conventional route, including but not limited to orally,parenterally, rectally or by inhalation (e.g. in the form of aerosols).In one embodiment, the compound or composition is administered by an IVinjection.

The pharmaceutical compositions for parenteral administration can beemulsions or homogeneous solutions. Use may be made, as solvent orvehicle, of propylene glycol, a polyethylene glycol, vegetable oils, inparticular olive oil, or injectable organic esters, for example ethyloleate. These pharmaceutical compositions can also contain adjuvants, inparticular wetting, isotonizing, emulsifying, dispersing and stabilizingagents. Sterilization can be carried out in several ways, for exampleusing a 0.2μ filter, by radiation or by heating (see, Remington'sPharmaceutical Sciences, 21st ed., Mack Publishing, Easton Pa. (2005).They can also be prepared in the form of sterile solid pharmaceuticalcompositions which can be dissolved at the time of use in sterile wateror any other injectable sterile medium.

The pharmaceutical compositions can also be aerosols. For use in theform of liquid aerosols, the pharmaceutical compositions can be sterilesolutions or solid pharmaceutical compositions dissolved at the time ofuse in apyrogenic sterile water, in saline or any other pharmaceuticallyacceptable vehicle. For use in the form of dry aerosols intended to bedirectly inhaled, the active principle is finely divided and combinedwith a water-soluble solid diluent or vehicle, for example dextran,mannitol or lactose.

Pharmaceutical compositions can be used in the preparation ofindividual, single unit dosage forms. Pharmaceutical compositions anddosage forms comprise compound and one or more excipients.

Pharmaceutical compositions and dosage forms can also comprise one ormore additional active ingredients. Examples of optional second, oradditional, active ingredients are disclosed herein.

In certain embodiments, the pharmaceutical composition provided hereinis a a single unit dosage form. Pharmaceutical compositions and singleunit dosage forms provided herein comprise a prophylactically ortherapeutically effective amount of compound or composition, andtypically one or more pharmaceutically acceptable carriers orexcipients. The term “carrier” refers to a diluent, adjuvant (e.g.,Freund's adjuvant (complete and incomplete)), excipient, or vehicle withwhich the therapeutic is administered. Such pharmaceutical carriers canbe sterile liquids, such as water and oils, including those ofpetroleum, animal, vegetable or synthetic origin, such as peanut oil,soybean oil, mineral oil, sesame oil and the like. In certainembodiments, water is a carrier when the pharmaceutical composition isadministered intravenously. Saline solutions and aqueous dextrose andglycerol solutions can also be employed as liquid carriers, particularlyfor injectable solutions. Examples of suitable pharmaceutical carriersare described in “Remington's Pharmaceutical Sciences” by E. W. Martin.

Typical pharmaceutical compositions and dosage forms comprise one ormore excipients. Suitable excipients are well-known to those skilled inthe art of pharmacy, and non limiting examples of suitable excipientsinclude starch, glucose, lactose, sucrose, gelatin, malt, rice, flour,chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodiumchloride, dried skim milk, glycerol, propylene, glycol, water, ethanoland the like. Whether a particular excipient is suitable forincorporation into a pharmaceutical composition or dosage form dependson a variety of factors well known in the art including, but not limitedto, the way in which the dosage form will be administered to a subjectand the specific active ingredients in the dosage form. Thepharmaceutical composition or single unit dosage form, if desired, canalso contain minor amounts of wetting or emulsifying agents, or pHbuffering agents.

Further provided herein are pharmaceutical compositions and dosage formsthat comprise one or more compounds that reduce the rate by which anactive ingredient will decompose. Such compounds, which are referred toherein as “stabilizers,” include, but are not limited to, antioxidantssuch as ascorbic acid, pH buffers, or salt buffers.

The pharmaceutical compositions and single unit dosage forms can takethe form of solutions, suspensions, emulsion, powders and the like. Suchcompositions and dosage forms will contain a prophylactically ortherapeutically effective amount of a prophylactic or therapeutic agent,in certain embodiments, in purified form, together with a suitableamount of carrier so as to provide the form for proper administration tothe subject. The formulation should suit the mode of administration. Inone embodiment, the pharmaceutical compositions or single unit dosageforms are sterile and in suitable form for administration to a subject,such as an animal subject, or a mammalian subject, and such as a humansubject.

A pharmaceutical composition provided herein is formulated to becompatible with its intended route of administration. Examples of routesof administration include, but are not limited to, parenteral, e.g.,intravenous, intradermal, subcutaneous, intramuscular, inhalation,intranasal, transdermal, topical, transmucosal, intra-tumoral,intra-synovial and rectal administration. In a specific embodiment, thecomposition is formulated in accordance with routine procedures as apharmaceutical composition adapted for intravenous, subcutaneous,intramuscular, intranasal or topical administration to human beings. Incertain embodiments, a pharmaceutical composition is formulated inaccordance with routine procedures for subcutaneous administration tohuman beings. In one embodiment, pharmaceutical compositions forintravenous administration are solutions in sterile isotonic aqueousbuffer. Where necessary, the pharmaceutical composition may also includea solubilizing agent and a local anesthetic such as lignocamne to easepain at the site of the injection.

Examples of dosage forms include, but are not limited to: liquid dosageforms suitable for parenteral administration to a subject; and sterilesolids (e.g., crystalline or amorphous solids) that can be reconstitutedto provide liquid dosage forms suitable for parenteral administration toa subject, including lyophilized solids.

The pharmaceutical composition, shape, and type of dosage forms providedherein will typically vary depending on their use. For example, a dosageform used in the initial treatment of disease may contain larger amountsof one or more of the active ingredients it comprises than a dosage formused in the maintenance treatment of the same infection. Similarly, aparenteral dosage form may contain smaller amounts of one or more of theactive ingredients it comprises than an oral dosage form used to treatthe same disease or disorder. These and other ways in which specificdosage forms encompassed herein will vary from one another will bereadily apparent to those skilled in the art. See, e.g., Remington'sPharmaceutical Sciences, 21st ed., Mack Publishing, Easton Pa. (2005).

Generally, the ingredients of pharmaceutical compositions providedherein are supplied either separately or mixed together in unit dosageform, for example, as a dry lyophilized powder or water free concentratein a hermetically sealed container such as an ampoule or sachetteindicating the quantity of active agent. Where the pharmaceuticalcomposition is to be administered by infusion, it can be dispensed withan infusion bottle containing sterile pharmaceutical grade water orsaline. Where the pharmaceutical composition is administered byinjection, an ampoule of sterile water for injection or saline can beprovided so that the ingredients may be mixed prior to administration.

In one embodiment, dosage forms provided herein comprise compound orcomposition within the range of about 1 mg/m² to about 150 mg/m² orabout 1 mg/m² to about 75 mg/m² per day, or per week, given as a singleonce-a-day dose in the morning or as divided doses throughout the daytaken with food. Particular dosage forms provided herein have about 1,3, 6, 9, 12, 15, 18, 21, 24, 27 or 30 mg/m² of the compound orcomposition.

5.7.1 Parenteral Dosage Forms

Parenteral dosage forms can be administered to patients by variousroutes including, but not limited to, subcutaneous, intravenous(including bolus injection), intramuscular, and intraarterial. Becausetheir administration typically bypasses patients' natural defensesagainst contaminants, parenteral dosage forms are sterile or capable ofbeing sterilized prior to administration to a patient. Examples ofparenteral dosage forms include, but are not limited to, solutions readyfor injection, dry products ready to be dissolved or suspended in apharmaceutically acceptable vehicle for injection, suspensions ready forinjection, and emulsions.

Suitable vehicles that can be used to provide parenteral dosage formsare well known to those skilled in the art. Examples include, but arenot limited to: Water for Injection USP; aqueous vehicles such as, butnot limited to, Sodium Chloride Injection, Ringer's Injection, DextroseInjection, Dextrose and Sodium Chloride Injection, and Lactated Ringer'sInjection; water-miscible vehicles such as, but not limited to, ethylalcohol, polyethylene glycol, and polypropylene glycol; and non-aqueousvehicles such as, but not limited to, corn oil, cottonseed oil, peanutoil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

Compounds that increase the solubility of one or more of the activeingredients disclosed herein can also be incorporated into theparenteral dosage forms. For example, cyclodextrin and its derivativescan be used to increase the solubility of active ingredients. See, e.g.,U.S. Pat. No. 5,134,127, which is incorporated herein by reference.

5.7.2 Topical and Mucosal Dosage Forms

In certain embodiments, provided herein are transdermal, topical, andmucosal dosage forms. Transdermal, topical, and mucosal dosage formsprovided herein include, but are not limited to, ophthalmic solutions,sprays, aerosols, creams, lotions, ointments, gels, solutions,emulsions, suspensions, or other forms known to one of skill in the art.See, e.g., Remington's Pharmaceutical Sciences, 21st ed., MackPublishing, Easton Pa. (2005); and Introduction to Pharmaceutical DosageForms, 5th ed., Lea & Febiger, Philadelphia (1990). Dosage formssuitable for treating mucosal tissues within the oral cavity can beformulated as mouthwashes or as oral gels. Further, transdermal dosageforms include “reservoir type” or “matrix type” patches, which can beapplied to the skin and worn for a specific period of time to permit thepenetration of a desired amount of active ingredients.

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide topical and mucosal dosage forms encompassedherein are well known to those skilled in the pharmaceutical arts, anddepend on the particular tissue to which a given pharmaceuticalcomposition or dosage form will be applied. With that fact in mind,typical excipients include, but are not limited to, water, acetone,ethanol, ethylene glycol, propylene glycol, butane-1,3-diol, isopropylmyristate, isopropyl palmitate, mineral oil, and mixtures thereof toform solutions, emulsions or gels, which are non-toxic andpharmaceutically acceptable. Moisturizers or humectants can also beadded to pharmaceutical compositions and dosage forms if desired.Examples of such additional ingredients are well known in the art. See,Remington's Pharmaceutical Sciences, 21st ed., Mack Publishing, EastonPa. (2005).

The pH of a pharmaceutical composition or dosage form may also beadjusted to improve delivery of one or more active ingredients.Similarly, the polarity of a solvent carrier, its ionic strength, ortonicity can be adjusted to improve delivery. Compounds such asstearates can also be added to pharmaceutical compositions or dosageforms to advantageously alter the hydrophilicity or lipophilicity of oneor more active ingredients so as to improve delivery. In this regard,stearates can serve as a lipid vehicle for the formulation, as anemulsifying agent or surfactant, and as a delivery-enhancing orpenetration-enhancing agent. Different salts, hydrates or solvates ofthe active ingredients can be used to further adjust the properties ofthe resulting pharmaceutical composition.

5.8 Analytical Methods

5.8.1 Analysis of Compositions Provided Herein

The amount of N-desmethyl-SNS-595 in compositions comprising SNS-595 andN-desmethyl SNS-595, including compositions provided herein can beestimated by methods known to one of skill in the art, such as HPLCanalysis. In an exemplary HPLC method, the mobile phase iswater/acetonitrile (0.1% TFA) gradient, eluting from 10% to 80%acetonitrile over 30 minutes with a gradient hold from 7 to 13 minutes(26% acetonitrile). The detection is performed at 275 nm on a reversephase column (C8).

FIG. 7 provides an HPLC chromatogram of an exemplary compositioncomprising about 99.2% SNS-595 and about 0.6% N-desmethyl-SNS-595.

6 EXAMPLES

Certain embodiments of the claimed subject matter are illustrated by thefollowing non-limiting examples.

Example 1 Preparation of an Exemplary Composition

A composition comprising SNS 595 and N-desmethyl-SNS-595 was preparedfrom (3S,4S)-3-methoxy-4-methylaminopyrrolidine di-p-toluenesulfonicacid andethyl-7-chloro-1,4-dihydro-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylate.An exemplary method for preparing starting materials is provided in FIG.2. Trans-3-benzylamino-1-tert-butyloxycarbonyl-4-hydroxypyrrolidine usedin the preparation of (3S,4S)-3-methoxy-4-methylaminopyrrolidinedi-p-toluenesulfonic acid was prepared as follows.

Preparation oftrans-3-benzylamino-1-tert-butyloxycarbonyl-4-hydroxypyrrolidine

Commercially available diallylamine was reacted with di-t-butyldicarbonate in heptane. After aqueous workup, the organic layer wasfurther diluted with heptanes (10 vol) and azeotropically distilled toremove residual water azeotropically distilled to a known volume and thesubsequent ring closing metathesis (RCM) reaction was carried byaddition of Grubbs catalyst (bis(tricyclohexylphosphine)benzylidineruthenium(IV) dichloride) in portions (0.1 mol % each, 2-3 portions)every two hours. The progress of the reaction was monitored by HPLCanalysis. The mixture was quenched with 0.02 N HCL (2×5 vol). The layerwas further washed with bicarbonate and water.

The ruthenium catalyst was deactivated by silica bound metal scavengers(Si-Thiol). The heptane solution containing the metathesis product 7 wasconcentrated to remove most of the solvent and a solvent-swap toacetonitrile (ACN) was implementated. Intermediate 7 was dissolved in 3volumes of ACN to a cold slurry (0° C.) of dibromo dimethyl hydantoin(DBH) in 3 volumes of ACN and 1 volume of water. After reactioncompletion, the mixture was quenched with 10% aq. Sodium thiosulfatesolution (3 volume) and extracted with ethyl acetate. The organic layerwas concentrated to remove most of the solvent. Resulting bromohydrinwas treated with 2 N NaOH (8 vol) to form the epoxide at ambienttemperature.

The epoxide was converted to racemic 9 by treatment with benzylamine(2.5 eq) with heating to 65° C. for 14 hours. The racemic intermediate 9precipitated out of the solution as it formed and was isolated byfiltration. The crude material was reslurried in cold MTBE (10° C.). Theresulting product was >99% pure.

Preparation of Ethyl1,4-Dihydro-7-[(3S,4S)-3-methoxy-4-methylamino-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylate

To a glass reactor covered with silver foil and equipped with anoverhead stirrer was added (3S,4S)-3-methoxy-4-methylaminopyrrolidinedi-p-toluenesulfonic acid (1190 g) and acetonitrile (9.0 L), and themixture stirred. The solution was cooled and triethylamine (TEA) (1560mL) was added, maintaining the batch temperature below 5° C.Ethyl-7-chloro-1,4-dihydro-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylate(750 g) was added, the ice-bath was removed and the reaction temperatureallowed to rise and then stirred an additional 48 hours. The reactionwas filtered, the filter cake was washed with acetonitrile (3×800 mL),and dried in vacuo to afford 936 g of the title compound as a whitesolid (97.6%).

Before working up the reaction, HPLC analysis of the reaction mixture(as described in section 5.4.1) was conducted to ensure thatintermediateethyl-7-chloro-1,4-dihydro-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylatewas <0.7%.

Preparation of a Composition Comprising1,4-Dihydro-7-[(3S,4S)-3-methoxy-4-methylamino-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylateand N-desmethyl1,4-Dihydro-7-[(3S,4S)-3-methoxy-4-methylamino-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylate

To a glass reactor covered with silver foil and equipped with anoverhead stirrer and a heating mantel was added ethyl1,4-Dihydro-7-[(3S,4S)-3-methoxy-4-methylamino-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylate(936 g), 1 N aqueous NaOH (3463) mL) and EtOH (112 mL) and thesuspension stirred. The reaction was adjusted to pH 7.5 with 25% aceticacid (660 mL), heated to 60° C. for approximately 2 hours, and thenallowed to cool. The resulting solids were filtered, washed with water(2×1498 mL) and EtOH (3×1498 mL), and dried in vacuo. The product wastransferred to a glass reactor equipped with an overhead stirrer,heating mantle and water filled condenser, and then diluted with EtOH(anhydrous; 16.8 L). The suspension was heated to 80° C. forapproximately 3 hours, allowed to cool, and the solids filtered, washedwith EtOH (anhydrous; 3×1498 mL) and dried in vacuo to afford 726 g ofthe composition comprising 99.2%1,4-Dihydro-7-[(3S,4S)-3-methoxy-4-methylamino-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylate(SNS 595) and 0.8% N-desmethyl1,4-dihydro-7-[(3S,4S)-3-methoxy-4-methylamino-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylate(N-desmethyl-SNS-595) as white solid (86.3%).

Example 2 Preparation and Characterization of a Hydrate of SNS-595

Methods of Preparation

The starting material, ethyl1,4-dihydro-7-[(3S,4S)-3-methoxy-4-methylamino-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylate,was prepared according to the reaction scheme depicted in FIG. 2. To aglass reactor covered with silver foil and equipped with an overheadstirrer and a heating mantel was added ethyl1,4-dihydro-7-[(3S,4S)-3-methoxy-4-methylamino-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylate(2685 g), 1 N aqueous NaOH (9935 mL) and EtOH (180 mL), and theresulting suspension was stirred. The reaction was adjusted to pH 7.5with 25% acetic acid (660 mL), heated to 60° C. for approximately 2hours, and then allowed to cool. The resulting solids were filtered,washed with water (2×4296 mL) and EtOH (3×4296 mL), and dried in vacuoat ambient temperature to give SNS-595 hydrate (2054 g).

Alternative Methods of Preparation

Other methods of preparation of this hydrate of SNS-595 may include,e.g.: dissolution of SNS-595 in water or an organic/water solvent systemfollowed by precipitation of this hydrate of SNS-595; slurry of a solidform of SNS-595 in water or an organic/water solvent system followed bytransformation to this hydrate of SNS-595; and exposure of a solid formof SNS-595 to humidity, heat and/or reduced pressure, thereby formingthis hydrate of SNS-595.

Methods of Characterization

X-Ray Powder Diffraction (XRPD) Analysis

XRPD patterns were obtained on a Scintag X₂ θ/θ diffractometer operatingwith copper radiation at 45 kV and 40 mA, using a Thermo ARLPeltier-cooled solid-state detector. Source slits of 2 and 4 mm, anddetector slits of 0.5 and 0.3 mm, were used for data collection.Unmilled material was placed in a stainless steel sample holder, leveledusing a glass microscope slide, and scanned on a six-positionauto-sampler. Powder diffraction patterns of the samples were obtainedfrom 2° to 42° 2θ at 1°/min. Calibration of the diffractometer wasverified using a silicon powder standard. Raw data files were convertedto ACS II format, transferred to a computer and displayed in Origin® 6.1for Windows.

Differential Scanning Calorimetry (DSC) Analysis

DSC measurements were collected using a PerkinElmer Pyris 1 DSC systemequipped with an Intracooler 2P refrigeration unit. The Pyris 1 DSC waspurged with nitrogen. Calibration was performed prior to analysis usingan Indium standard at a 10° C./min heating rate. Approximately 2 mgsamples were individually placed in tared PerkinElmer 25-μL universalaluminum pans with holes in the lids. The samples were weighed on aSartorius microbalance and sealed using a PerkinElmer pan crimper press.Samples were heated from approximately 25° C. to approximately 350° C.at approximately 10° C./min.

Thermogravimetric Analysis (TGA)

TGA measurements were collected using a PerkinElrner TGA 7 purged withnitrogen. A 100-mg standard weight and nickel sample were used to verifybalance and temperature calibrations, respectively. Samples were heatedfrom approximately 25° C. to approximately 350° C. at approximately 10°C./min.

Water Content Analysis

The amount of water in a hydrate may be analyzed using a number oftechniques as understood in the art. For example, the amount of watermay be determined based on the observed weight loss in a TGA thermogram.In addition, the exhaust from a TGA furnace may be coupled to aninstrument of chemical analysis, such as an mass spectrometry instrumentor an infrared spectroscopy instrument, to confirm the chemical purityof the water vapor emitted upon heating. Moreover, Karl Fischer (KF)analysis may be used to analyze the water content of a hydrate sample.Coulometric KF analysis for water determination may be performed using aMettler Toledo DL39 Karl Fischer titrator. Approximately 14-32 mg of asample is placed in the KF titration vessel containingHYDRANAL®-Coulomat AD reagent for coulometric KF titration and mixed for60 seconds to ensure dissolution. The sample is then titrated by meansof a generator electrode which produces iodine by electrochemicaloxidation. The analysis is repeated to ensure reproducibility of themeasurements.

Characterization

A hydrate of SNS-595 was characterized by X-ray powder diffraction(XRPD) using the methods described above. A representative XRPD patternof this hydrate of SNS-595 is shown in FIG. 8. The 2θ angles (Cu Kαradiation), D-spacing values and relative intensities for certain XRPDpeaks of this hydrate of SNS-595 are provided in Table 1.

TABLE 1 Approximate Peak Approximate Peak Relative Peak IntensityPosition Position (as percentage of maximum (in Degrees 2θ) (inD-spacing in Å) peak intensity) 6.8913 12.8164 15.21 8.1556 10.8321100.00 11.1325 7.9413 11.17 16.3550 5.4154 9.45 17.5225 5.0571 7.7518.8469 4.7046 11.84 20.7694 4.2733 9.15 24.9131 3.5711 6.70

The hydrate of SNS-595 was also characterized by thermogravimetricanalysis and differential scanning calorimetry, using the methodsdescribed above. Representative TGA and DSC thermograms are shown inFIG. 9. The TGA thermogram exhibited a weight loss of approximately4.44% between the temperatures of approximately 25° C. and approximately200° C. The DSC thermogram exhibited an endothermic event with an onsettemperature of approximately 126.5° C., a peak temperature ofapproximately 138.8° C., and a heat of fusion of approximately 85.1 J/g.The DSC thermogram also exhibited an endothermic event with an onsettemperature of approximately 273.3° C.

Total water content in particular samples of this hydrate of SNS-595include values ranging between: 0.8 to 1.2 molar equivalents of waterper mole of SNS-595; 0.9 to 1.1 molar equivalents of water per mole ofSNS-595; 0.95 to 1.05 molar equivalents of water per mole of SNS-595;and 0.98 to 1.02 molar equivalents of water per mole of SNS-595.

Example 3 Pharmaceutical Composition Suitable for Injection orIntravenous Infusion

An illustrative example of a suitable pharmaceutical compositioncomprises: 10 mg total SNS-595 and N-desmethyl-SNS-595 per mL of aqueoussolution of 4.5% sorbitol that is adjusted to pH 2.5 withmethanesulfonic acid. One protocol for making such a solution includesthe following for making a 100 mg/10 mL presentation: 100 mg totalSNS-595 and N-desmethyl-SNS-595 (about 99.2 mg SNS-595 and about 0.8 mgN-desmethyl-SNS-595) and 450 mg D-sorbitol are added to distilled water;the volume is brought up to a volume of 10 mL; and the pH of theresulting solution is adjusted to 2.5 with methanesulfonic acid. Theresulting composition is also suitable for lyophilization. Thelyophilized form is then reconstituted with sterile water to theappropriate concentration prior to use.

Example 4 Pharmaceutical Composition Suitable for Injection orIntravenous Infusion

An illustrative example of a suitable pharmaceutical compositioncomprises: 10 mg total SNS-595 and N-desmethyl-SNS-595 per mL of aqueoussolution of 4.5% sorbitol that is adjusted to pH 2.5 withmethanesulfonic acid. One protocol for making such a solution includesthe following for making a 100 mg/0 mL presentation: 100 mg totalSNS-595 and N-desmethyl-SNS-595 (about 98.5 mg SNS-595 and about 1.5 mgN-desmethyl-SNS-595) and 450 mg D-sorbitol are added to distilled water;the volume is brought up to a volume of 10 mL; and the pH of theresulting solution is adjusted to 2.5 with methanesulfonic acid. Theresulting composition is also suitable for lyophilization. Thelyophilized form is then reconstituted with sterile water to theappropriate concentration prior to use.

Example 5 Comparative cytotoxicity of SNS-595, N-desmethyl-SNS-595 andO-desmethyl-SNS-595 in MTT Assay

Preparation of Samples

Stock Solutions

MTT Dye Solution: 5 mg/mL MTT dye (Sigma #M2128) in sterile PBS, filtersterilized, stored at 4° C. in a dark bottle no longer than 1 month.

Lysis Buffer: 50% N,N-dimethyl formamide (Aldich #31, 993-7) in dH₂O,20% w/v lauryl sulfate (added with stirring at 37° C.) (Sigma #L4509),adjusted pH to 4.7 with 2.5% of 80% acetic acid (Fisher #A490-212) and2.5% 1 NHC1 (Spectrum #HY105).

Supplemented RPMI media: 500 mL RPMI (Cellgro #10-040-CV), 10% FBS(Cellgro #35-01 lCV) and 1% Sodium Bicarbonate solution (Cellgro@35-035-CI), 1% Antibotic Solution (100× stock; Cellgro #30-004CI).

Trypsin (Cellgro #25-053-CL)

Cells

HCT-116 cells were purchased from ATTC (#CCL-247).

Controls

Background: No cells, Totals: Cells treated with DMSO only

Method

96-well tissue culture-treated flat bottom plates (Costar #3595) wereplated with 4000 trypsinized cells in 100 μL/well and incubatedovernight. 100× stock concentrations of compounds were prepared in DMSOand serially diluted two-fold in DMSO in a 96-well polypropylenev-bottom plate (Costar #3363). 5 μL of DMSO dilutions were then added to45 μL supplemented RPMI, and 10 μL/well of this mixture were added toplates containing HCT-116 cells. The highest concentration of compoundswas 5 μM. Plates were incubated for 72 hours at 37° C. in an incubator(5% CO₂).

After incubation, 20 μL of MTT solution were added to each well andincubated at 37° C. for 1.5 hour. 100 μL lysis buffer was then added andplates were incubated for 48 hours at 37° C. Cell viability was thenrecorded by absorbance at 595 nm using a Biorad Benchmark MicroplateReader. The fraction of dead cells was determined by

Fraction of Dead cells=Absorbance of sample well−Avg(no cellcontrol)/Avg(Absorbance of DMSO only Control)−Avg(no cell control).

The results of the cell viability assay are shown in FIG. 1. SNS-595reduced cell viability with an IC₅₀=425 nM in this assay.N-desmethyl-SNS-595 has an IC₅₀=507 nM. O-desmethyl-SNS-595 has anIC₅₀>>5 μM.

Example 7 Identification of N-desmethyl SNS-595

The in vitro production of N-Desmethyl SNS-595 from SNS-595 wasmonitored as described herein. The pooled Human Liver Microsomes wereobtained from BD Gentest and NADPH was obtained from Sigma.

The in vitro reactions were performed (a) in the presence of NADPH as acofactor and (b) in the absence of NADPH to identify any SNS-595derivatives generated by cofactor independent mechanisms.

The reaction mixture constituted 100 mM Sodium Phosphate buffer, pH 7.4,containing 3.3 mM MgCl₂, 1 mg/mL liver microsomal protein, 10 or 100 μMSNS-595, and 1 mM NADPH for reaction with NADPH. In both the NADPH plusand minus reactions, SNS-595 was added to the reaction mixture andallowed to equilibrate to 37° C. for 10 minutes before starting thereaction by adding NADPH or an equivalent volume of buffer. Reactionswere performed in a volume of 1 mL in 2 mL 96-well polypropylene assayblocks. The reaction was stopped by adding an equal volume ofacetonitrile at both 0 and 60 minutes after initiating the reaction.Samples were placed on ice until centrifugation at 4100 g for 10 minutesto remove proteins for subsequent analysis.

Analysis

SNS-595 and the reaction products were identified on an API 4000 massspectrometer coupled to a turbo electro spray ionization source. Priorto MS analysis chromatography was performed to achieve separation of thereaction precuts (N-desmethyl-SNS-595 had the same retention time asSNS-595). The HPLC system consisted of an Agilent 1100 binary pump andsingle wavelength UVN is as a detector, and a Phenomenex SynergiHydro-RP column (150×2 mm, 4 micron, 80 Å particle size). Followingcolumn separation, UV spectra were collected at 350 nm before massspectral analysis. Mass spectral identification of the reaction productswas done using a series of three experiments; MS full scan, product ionscans on all reaction products identified in the MS full scan analysisas well as multiple reaction monitoring (MRM) or product ion scans ontheoretically possible reaction products. Product ion scans wereperformed at collision energies of 50 volts for SNS-595 and the reactionproducts.

N-desmethyl SNS-595 and O-desmethyl SNS-595 were synthesized. Retentiontime and fragmentation pattern of the authentic standards was comparedwith those of the reaction product peaks and used to verify the identityof the products of the in vitro reaction.

Results

Identification of the N-desmethyl SNS-595 was based on a comparison ofretention time and fragmentation patterns with those of the chemicallysynthesized N-desmethyl reference standard. As shown in FIG. 3,N-desmethyl-SNS-595 (peak 2) is the predominant product formed. At a 10μM incubation concentration, N-desmethyl-SNS-595 is the only productdetectable. The second product formed after incubation of highconcentrations of SNS-595 was identified as O-desmethyl-SNS-595 (peak 1in FIG. 3). N-desmethyl SNS-595 has a retention time of 5.33 min and ischromatographically separated from peak 1 in both chromatograms in FIG.3 (4.79 min; small peak seen in pane A not visible in pane B at thescale shown).

Chromatographic identification of peak 2 as N-desmethyl SNS-595(retention time 5.33 min) is shown in FIG. 4. The retention time ofcompounds corresponding to peak 2 is the same as the chemicallysynthesized standard of N-desmethyl SNS-595, shown in pane B. Peak 2 isalso resolved from the O-desmethyl SNS-595 which has a retention time4.79 min. Confirmation of O-desmethyl SNS-595 was made throughcomparison with a chemically synthesized reference compound.

Mass spectra identification of peak 1 and 2 as demethylated products ofSNS-595 was made by comparison between the product ion spectra ofSNS-595 and peaks 1 and 2. FIG. 5 shows the product ion spectra ofSNS-595 with structures assigned to the predominant m/z fragments. InFIG. 5, structure A is SNS-595 and structures B and C are assigned tofragmentation on the carboxylic acid region of the molecule having amass loss of 18 and 70 amu, respectively. Structure D is assigned to them/z 269 fragment, which does not contain the 3-methoxy or 4-methylaminomoieties of the pyrrolidinyl ring. Structure E also lacks the 3-methoxyand 4-methylamino pieces and is assigned to m/z 243.

FIG. 6 is the spectra from peak 2 with structures A-E assigned to themajor fragments (spectra from the authentic reference N-desmethylSNS-595 standard is identical). Structure A and fragments B, and C showthe same delta mass loss as fragments B and C in the SNS-595 spectra,indicating peak 2 is structurally similar to SNS-595. In addition,structures A, B, and C have a mass of 14 amu less than the correspondingfragments from the SNS-595 spectra indicating a loss of a methyl group.Structures D and E share the same mass fragments in both the spectra forpeak 2 and SNS-595 showing the loss of 14 amu results fromN-demethylation.

The embodiments of the claimed subject matter described above areintended to be merely exemplary, and those skilled in the art willrecognize, or will be able to ascertain using no more than routineexperimentation, numerous equivalents of specific compounds, materials,and procedures. All such equivalents are considered to be within thescope of the claimed subject matter and are encompassed by the appendedclaims.

1. A composition comprising about 97% to about 99.99% by weight of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid and from about 0.01% up to about 3% by weight of an N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid, wherein each of the percentage is based upon total weight of thetwo components.
 2. The composition of claim 1, comprising about 0.8% byweight N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid.
 3. The composition of claim 1, comprising about 1.5% by weightN-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid.
 4. The composition of claim 1, comprising about 98.5% by weight(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid.
 5. The composition of claim 1, comprising about 99.2% by weight(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid.
 6. A hydrate of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid, wherein the hydrate exhibits an X-ray powder diffraction patterncomprising a peak at approximately 8.2 degrees 2θ.
 7. The hydrate ofclaim 6 that exhibits an X-ray powder diffraction pattern furthercomprising peaks at approximately 6.9, 11.1 and 18.8 degrees 2θ.
 8. Thehydrate of claim 7 that exhibits an X-ray powder diffraction patternfurther comprising peaks at approximately 16.4, 17.5, 20.8 and 24.9degrees 2θ.
 9. A hydrate of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid, wherein the hydrate exhibits a differential scanning calorimetrythermogram comprising an endothermic event with an onset temperature ofapproximately 126.5° C. when heated from approximately 25° C. toapproximately 350° C. at approximately 10° C./min.
 10. The hydrate ofclaim 9 that exhibits a differential scanning calorimetry thermogramfurther comprising an endothermic event with an onset temperature ofapproximately 273.3° C.
 11. A hydrate of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid, wherein the hydrate exhibits a thermogravimetric analysisthermogram comprising a weight loss of approximately 4.4% when heatedfrom approximately 25° C. to approximately 200° C. at approximately 10°C./min.
 12. A hydrate of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid, wherein the hydrate comprises between approximately 0.8 and 1.2molar equivalents of water per mole of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid.
 13. The hydrate of claim 12 that comprises between approximately0.9 to 1.1 molar equivalents of water per mole of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid.
 14. The hydrate of claim 12 that comprises between approximately0.95 to 1.05 molar equivalents of water per mole of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid.
 15. The hydrate of claim 12 that comprises between approximately0.98 to 1.02 molar equivalents of water per mole of(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid.
 16. A pharmaceutical composition comprising the composition ofclaim 1 and a pharmaceutically acceptable carrier, excipient oradjuvant.
 17. A pharmaceutical composition comprising the hydrate ofclaim 6 and a pharmaceutically acceptable carrier, excipient oradjuvant.
 18. A method for treating cancer comprising administering thecomposition of claim 1 to a mammal in need of thereof.
 19. A method fortreating cancer comprising administering the hydrate of claim 6 to amammal in need of thereof.
 20. The method of claim 18, wherein thecomposition is administered in a dose of about 1 mg/m²-150 mg/m² to amammal in need of thereof.
 21. A method for treating cancer comprisingadministering a N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid in a dose of about 1 mg/m²-150 mg/m² to a mammal in need ofthereof.
 22. The method of claim 18, wherein the dose is about 15 mg/m².23. The method of claim 18, wherein the dose is about 48 mg/m².
 24. Themethod of claim 18, wherein the dose is about 60 mg/m².
 25. The methodof claim 18, wherein the dose is about 75 mg/m².
 26. The method of claim18, wherein the cancer comprises a solid tumor.
 27. The method of claim18, wherein the cancer is a bladder cancer, breast cancer, cervicalcancer, colon cancer, esophageal cancer, head and neck cancer, livercancer, lung cancer, melanoma, mycloma, neuroblastoma, ovarian cancer,pancreatic cancer, prostate cancer, renal cancer, sarcoma, skin cancer,stomach cancer, testicular cancer, thyroid cancer, uterine cancer orhematologic malignancy.
 28. The method of claim 18, wherein the canceris a colon cancer, pancreas cancer, breast cancer, mesothelioma,cholangiocarcinoma, leiomyosarcoma, liposarcoma, melanoma,nasopharyngeal cancer, neuroendocrine cancer, ovarian cancer, renalcancer, salivary gland cancer, small cell lung cancer, leukemia orspindle cell carcinoma.
 29. The method of claim 18 further comprisingadministering a therapeutically effective dose of a second agent.
 30. Amethod for treating cancer comprising: i) administering a dose of about1 mg/m²-150 mg/m² of the composition of claim 1 to a mammal; ii) waitinga period of at least 2 days where the mammal is not administered thecomposition; and iii) administering another dose of 1 mg/m²-150 mg/m² ofthe composition to the mammal.
 31. A method for treating cancercomprising: i) administering a dose of about 1 mg/m²-150 mg/m² of thecomposition of claim 1 to a mammal; ii) waiting a period of at least 7days where the mammal is not administered the composition; and iii)administering another dose of 1 mg/m²-150 mg/m² of the composition tothe mammal.
 32. The method of claim 30, wherein the dose of thecomposition is 1 mg/m²-75 mg/m².
 33. The method of claim 30, wherein thedose of the composition is 10 mg/m²-60 mg/m².
 34. The method of claim30, wherein the dose of the composition is 1 mg/m²-48 mg m².
 35. Themethod of claim 30, wherein the dose of the composition is 75 mg/m². 36.The method of claim 30, wherein the dose of the composition is 60 mg/m².37. The method of claim 30, wherein the dose of the composition is 48mg/m².
 38. The method of claim 30, wherein the dose of the compositionis 15 mg/m².
 39. A method for treating cancer comprising: i)administering a dose of about 1 mg/m²-150 mg/m² of N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to a mammal; ii) waiting a period of at least two days where themammal is not administered any N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid; and iii) administering another dose of 1 mg/m²-150 mg/m² ofN-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to the mammal.
 40. The method of claim 39, wherein the dose isabout 1 mg/m² to 75 mg/m².
 41. The method of claim 39, wherein the doseis about 75 mg/m².
 42. The method of claim 39, wherein the dose is about60 mg/m².
 43. The method of claim 39, wherein the dose is about 48mg/m².
 44. The method of claim 39, wherein the dose is about 15 mg/m².45. A method for treating cancer comprising administering a dose ofabout 15 mg/m² of the composition of claim 1 to a mammal once a week,wherein the one-week period comprises a treatment cycle and thetreatment cycle is repeated at least twice.
 46. A method for treatingcancer comprising administering a dose of about 48 mg/m² of thecomposition of claim 1 to a mammal once in a three week period, whereinthe three week period comprises a treatment cycle.
 47. A method fortreating cancer comprising administering a dose of about 60 mg/m² of thecomposition of claim 1 to a mammal once in a three week period, whereinthe three week period comprises a treatment cycle.
 48. A method fortreating cancer comprising administering a dose of about 15 mg/m² ofN-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to a mammal once a week, wherein the one-week period comprises atreatment cycle and the treatment cycle is repeated at least twice. 49.A method for treating cancer comprising administering a dose of about 48mg/m² of N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to a mammal once in a three week period, wherein the three weekperiod comprises a treatment cycle.
 50. A method for treating cancercomprising administering a dose of about 60 mg/m² of N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to a mammal once in a three week period wherein the three weekperiod comprises a treatment cycle.
 51. A method for treating cancercomprising: i) administering a dose of 1 mg/m²-75 mg/m² of N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to a mammal; ii) waiting a period of at least 2 days where themammal is not administered any N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid; and iii) administering another dose of 1 mg/m²-75 mg/m² ofN-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to the mammal.
 52. A method for treating cancer comprising: i)administering a dose of 3 mg/m²-24 mg/m² of N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to a mammal; ii) waiting a period of at least 7 days where themammal is not administered any N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid; and iii) administering another dose of 3 mg/m²-24 mg/m² ofN-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to the mammal.
 53. The method of claim 51, wherein the dose is 15mg/m², 48 mg/m², 60 mg/m² or 75 mg/m²
 54. A method for treating cancercomprising: i) administering a dose of 10 mg/m²-60 mg/m² of N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to a mammal; ii) waiting a period of at least 7 days where themammal is not administered any N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid; and iii) administering another dose of 10 mg/m²-60 mg/m² ofN-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to the mammal.
 55. The method of claim 39, wherein the dose is 15mg/m² and the waiting period is 14 days.
 56. The method of claim 55,wherein the waiting period is 28 days.
 57. A method for treating cancercomprising: i) administering a dose of 1 mg/m²-75 mg/m² of thecomposition of claim 1 once a week for three weeks to a mammal; ii)waiting a period of at least two days where the mammal is notadministered the composition; and iii) administering another dose of 1mg/m²-75 mg/m² of the composition to the mammal.
 58. The method of claim57, wherein the dose is 15 mg/m², 48 mg/m², 60 mg/m² or 75 mg/m².
 59. Amethod for treating cancer comprising: i) administering a dose of 3mg/m²-24 mg/m² of the composition of claim 1 once a week for three weeksto a mammal; ii) waiting a period of at least two days where the mammalis not administered the composition; and iii) administering another doseof 3 mg/m²-24 mg/m² of the composition to the mammal.
 60. A method fortreating cancer comprising: i) administering a dose of 10 mg/m²-60 mg/m²of the composition of claim 1 once a week for three weeks to a mammal;ii) waiting a period of at least two days where the mammal is notadministered the composition; and iii) administering another dose of 10mg/m²-60 mg/m² of the composition to the mammal.
 61. A method fortreating cancer comprising a treatment cycle, wherein the treatmentcycle comprises administering a dose of 3 mg/m²-24 mg/m² of thecomposition of claim 1 to a mammal once a week for three weeks followedby a period of at least two weeks where no composition is administeredto the mammal.
 62. A method for treating cancer comprising a treatmentcycle, wherein the treatment cycle comprises administering a dose of 10mg/m²-60 mg/m² of the composition of claim 1 to a mammal once a week forthree weeks followed by a period of at least two weeks where nocomposition is administered to the mammal.
 63. A method for treatingcancer comprising a treatment cycle, wherein the treatment cyclecomprises administering a dose of 3 mg/m²-24 mg/m² of the composition ofclaim 1 to a mammal on days 1, 4, 8 and 11 followed by a period of 28days where no composition is administered to the mammal.
 64. A methodfor treating cancer comprising a treatment cycle, wherein the treatmentcycle comprises administering a dose of 10 mg/m²-60 mg/m² of theN-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to a mammal once a week for three weeks followed by a period of atleast two weeks where no compound is administered to the mammal.
 65. Amethod for treating cancer comprising a treatment cycle, wherein thetreatment cycle comprises administering a dose of 3 mg/m²-24 mg/m² ofthe N-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to a mammal once a week for three weeks followed by a period of atleast two weeks where no compound is administered to the mammal.
 66. Themethod of claim 45, wherein the cycle is repeated a plurality of times.67. The method of claim 59, wherein the dose is 15 mg/m².
 68. A methodfor treating cancer comprising a treatment cycle, wherein the treatmentcycle comprises administering a dose of about 48 mg/m² or 60 mg/m² ofN-desmethyl(+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylicacid to a mammal once every three weeks.
 69. A method for treatingcancer comprising a treatment cycle, wherein the treatment cyclecomprises administering a dose of about 48 mg/m² or 60 mg/m² of thecomposition of claim 1 to a mammal once every three weeks.
 70. Themethod of claim 18, wherein the compound is administered as an IVinjection.
 71. The method of claim 18, wherein the composition isadministered as an IV injection.
 72. The method of claim 18, wherein thecancer is refractory.
 73. The method of claim 18, wherein the mammal isa human.